Lipid-based platinum compounds and nanoparticles

ABSTRACT

The present disclosure is in relation to the field of nanotechnology and cancer therapeutics. In particular, the present disclosure relates to platinum based compounds comprising platinum moiety, linker moiety and lipid moiety and corresponding nanoparticles thereof. The disclosure further relates to synthesis of said platinum based compounds, nanoparticles and compositions comprising said platinum based compounds/nanoparticles. The disclosure also relates to methods of managing cancer by employing aforesaid carbene compounds, platinum based compounds, nanoparticles and compositions thereof.

CROSS-REFERENCE TO RELATED APPLICATIONS

This application is a divisional of U.S. patent application Ser. No.16/123,430, filed Sep. 6, 2018, which is a divisional of U.S. patentapplication Ser. No. 14/898,355, filed Dec. 14, 2015, now U.S. Pat. No.10,081,648, issued Sep. 25, 2018, which is the national stage ofInternational Patent Application No. PCT/US14/42339, filed Jun. 13,2014, which claims the benefit of Indian Patent Application No.1781/DEL/2013, filed Jun. 14, 2013, the contents of which areincorporated herein by reference in their entireties.

TECHNICAL FIELD

The present disclosure is in relation to the field of nanotechnology andcancer therapeutics. In particular, the present disclosure relates toplatinum based compounds comprising platinum moiety, linker moiety andlipid moiety and corresponding nanoparticles thereof. The disclosurefurther relates to synthesis of said platinum based compounds,nanoparticles and compositions comprising said platinum basedcompounds/nanoparticles. The disclosure also relates to methods ofmanaging cancer by employing aforesaid platinum based compounds,nanoparticles and compositions.

BACKGROUND

The use of nanotechnology in cancer is emerging globally. Although thereare few reports on nanoparticles in cancer therapy but all have variousdrawbacks such as toxicity, low release kinetics of drug, lowcirculation stability and so on.

Lipidic nanoparticles (e.g. Doxil, a pegylated liposomal formulation ofdoxorubicin hydrochloride) and albumin-complexes (e.g. Abraxane, apaclitaxel-albumin complex) nanoparticles are used in humans and havebeen demonstrated as having improved systemic toxicity profile and havehelped resolve certain formulation challenges (Ferrari M, Nature Rev.Cancer, 2005, 5:161). Platinum-based chemotherapeutic agents are used asfirst line of therapy in over 70% of all cancers. Cisplatin undergoesrapid formation of cis-[Pt(NH₃)₂C(OH₂)]⁺ and cis-[Pt(NH₃)₂(OH₂)]²⁺resulting in nephrotoxicity. Further, aquation of both carboplatin andoxaliplatin are significantly slower, resulting in decreased potency. Inthe recent past, considerable progress has been made wherein, Dhar et al(PNAS, 2008, 105, 17356) generated a platinum (IV) complex(c,t,c-[Pt(NH₃)₂(O₂CCH₂CH₂CH₂CH₂CH₃)₂Cl₂] that is hydrophobic enough forencapsulation into PLGA-b-PEG nanoparticles. However, the prodrug inthis case has to be intracellularly processed into cisplatin.Furthermore, alternative strategies based on conjugation of platinum topolymers (e.g. a polyamidoamine dendrimer-platinum complex) resulted ina 200-550 fold reduction in cytotoxicity than free cisplatin. This was aresult of strong bonds formed between the polymer and platinum (J PharmSci, 2009, 98, 2299). Another example is AP5280, a N-(2-hydroxypropyl)methacrylamide copolymer-bound platinum that is less potent thancarboplatin. Here, the platinum is held by an aminomalonic acidchelating agent coupled to the COOH-terminal glycine of a tetrapeptidespacer (Clin Can Res, 2004, 10, 3386; Eur J Can, 2004, 40, 291).

Further, WO 2010/091192 A2 (Sengupta et al) discloses biocompatibleconjugated polymer nanoparticles including a copolymer backbone, aplurality of sidechains covalently linked to said backbone, and aplurality of platinum compounds dissociably linked to said backbone. Thedisclosure is further directed to dicarbonyl-lipid compounds wherein aplatinum compound is dissociably linked to the dicarbonyl compound.

However, various drawbacks are associated with the presently employednanoparticles. The present disclosure aims at overcoming the drawbacksof the prior art and providing for stable, potent and safernano-platinates in cancer chemotherapy.

SUMMARY

In one aspect, the disclosure provides a compound comprising: (a) aplatinum moiety; and (b) a lipid connected to said platinum moiety. Insome embodiments, the compound is of formula (VIII):

Q-linker-lipid  (VIII),

-   -   wherein:    -   Q is a platinum containing moiety and the linker has at least        one linkage to the platinum atom.

The disclosure also provides a method of obtaining Pt-lipid moleculesdisclosed herein. Accordingly, in one aspect the disclosure provides amethod of obtaining a compound comprising: (a) a platinum moiety, and alipid connected to said platinum moiety a method of obtaining a compoundcomprising, said method comprising conjugating the lipid with theplatinum moiety to obtain said compound.

The disclosure also provides particles, such as nanoparticles comprisingone or more of the Pt-lipid molecules disclosed herein. Thus, in oneaspect, the disclosure provides a particle, for example, but notlimited, a nanoparticle comprising a platinum based compound, whereinthe platinum based compound comprises: (a) a platinum moiety; and (b) alipid connected to said platinum moiety.

The disclosure also provides a pharmaceutical composition comprising thecompound as disclosed above or the nanoparticle as disclosed above or acombination thereof, along with pharmaceutically acceptable excipient;and a method of managing or treating cancer, said method comprising stepof administering the compound as disclosed above or the nanoparticle asdisclosed above or the composition as disclosed above, to a subject inneed thereof.

BRIEF DESCRIPTION OF THE DRAWINGS

In order that the invention may be readily understood and put intopractical effect, reference will now be made to exemplary embodiments asillustrated with reference to the accompanying figures. The figurestogether with a detailed description below, are incorporated in and formpart of the specification, and serve to further illustrate theembodiments and explain various principles and advantages, in accordancewith the present disclosure.

FIGS. 1A-1C depict the synthesis procedure of Cholesterol-Oxaliplatincompounds (Formula I) with carbamate linkage (Compounds 1, 2 and 3).Reagents and Conditions: a) Ethylenediamine (20 eq), Dry DCM, 0° C.-roomtemperature (RT), 24 hours; b) succinic anhydride, DCM, pyridine, RT, 24hours; c) malonic acid monoethyl ester, DCM, EDCl, HOBt, RT, 24 hours;d, d′) LiOH, THF, H₂O, 3 hours, RT; e) oxalic acid monoethyl ester, DCM,EDCl, HOBt, RT, 24 hours; f) AgNO₃, H₂O, RT, 24 hours; g, g′, g″) DMF,H₂O, RT, 24 hours.

FIGS. 2A-2C depict the synthesis procedure of Cholesterol-Oxaliplatincompounds (Formula I) with ether linkage (Compounds 6, 4 and 5).Reagents and Conditions: a) TsCl, Dry DCM, Pyridine, RT, 6 hours; b)ethylene glycol, dioxane, reflux, 4 hours; c) TsCl, DCM, Pyridine, RT, 6hours; d) NaN₃, DMF, 3 hours, RT; e) PPh₃, THF, H₂O, RT, 4 hours; f)succinic anhydride, DCM, pyridine, RT, 24 hours; g) malonic acidmonoethyl ester, DCM, EDCl, HOBt, RT, 24 hours; h,h′) LiOH, THF, H₂O, 3hours, RT; i) oxalic acid monoethyl ester, DCM, EDCl, HOBt, RT, 24hours; j) AgNO₃, H₂O, RT, 24 hours; k,k′,k″) DMF, H₂O, RT, 24 hours.

FIGS. 3A-3E depict the synthesis procedure of compounds of Formula III.FIG. 3A represents the synthesis of Compound 32 (wherein, R=Cholesterolor other lipid). FIG. 3B represents the synthesis of Compound 33(wherein, R=Cholesterol or other lipid). FIG. 3C represents thesynthesis of Compound 34 (wherein, R=Cholesterol or other lipid). FIG.3D represents the synthesis of Compound 35 (wherein, R=Cholesterol orother lipid). FIG. 3E represents the synthesis of Compound 36 (wherein,R=Cholesterol or other lipid);

FIGS. 4A-4E depict the synthesis procedure of compounds of Formula II.FIG. 4A represents the synthesis of Compound 38. FIG. 4B represents thesynthesis of Compound 39. FIG. 4C represents the synthesis of Compound40. FIG. 4D represents the synthesis of Compound 41. FIG. 4E representsthe synthesis of Compound 42.

FIG. 5 depicts the physicochemical characterization of nanoparticles.DLS plot is represented which show the distribution of particle size.

FIGS. 6A-6C depict the in-vitro characterization of synthesizedcholesterol-oxaliplatin nanoparticle compositions. The graphs show theconcentration-effect of different cholesterol-oxaliplatin nanoparticlecompositions and Oxaliplatin (control) on cellular viability of 4T1(breast cancer cell line) (FIG. 6A), HeLa (cervical cancer cell line)(FIG. 6B) and LLC (lung cancer cell line) (FIG. 6C) cancer cells asmeasured using MTS assay. The x-axis depicts the equivalentconcentrations of platinum.

FIGS. 7A-7F show MTT assay results for some exemplary compoundsdisclosed herein. Graphical representation of MTT assay for the activityof IO-compounds examined on various human cancer cell lines. The MTTassay showed a reduction in cell viability for the various exemplarycompounds tested. The IC₅₀ values of compound/oxaliplatin tested incell-lines are mentioned alongside, in corresponding colors as the cellviability curve.

FIG. 8 shows cellular uptake of platinum compounds. Cells were incubatedwith 50 μM concentration of platinum compounds for 5 hours. The amountof platinum accumulated in cells was measured by AAS and expressed as ngof platinum accumulated per 10⁵ cells.

FIG. 9 shows platinum distribution in tumors. The total platinum contentin tumors was measured by AAS and expressed as ng of platinumaccumulated per mg of tumor.

DETAILED DESCRIPTION

In some embodiments, platinum based compounds are disclosed whichcomprises: (a) a Platinum moiety; (b) at least one linker connected tosaid Platinum moiety; and (c) a lipid connected to said linker.

In the compounds disclosed herein, the platinum moiety is linked to thelipid molecule either directly or via a linker molecule. In someembodiments, the platinum moiety is linked to the lipid molecule via alinker molecule. For example, the presence of a linker can provide for acarbamate and/or ether linkage connecting a dicarbonyl molecule (forlinking with the platinum moiety) and the lipid molecule. In some otherembodiments of the present disclosure, the platinum moiety is directlyconnected to the lipid molecule. All possible linker molecules providinga carbamate and/or ether linkage form a part of the instant disclosure.

In some embodiments, the platinum based compound disclosed herein is acompound of Formula (VIII):

Q-linker-lipid  (VII),

-   -   wherein:    -   Q is a platinum containing moiety and the linker has at least        one linkage to the platinum atom.

In some embodiments of the various aspects described herein, Q is

wherein: X₃ is selected from a group comprising (CH₂)_(n), CH₂—NH andC₄H₈; X₄ is CO or —CH—CH₃; Z is a platinum containing compound, whereinthe platinum forms a part of the ring; and n is 0, 1, or 2.

In some embodiments of the various aspects described herein, Q is

wherein: X is NH or N(CH₂COO⁻); and Z is a platinum containing compound,wherein the platinum forms a part of the ring.

In some embodiments of the various aspects described herein, Q is

wherein: X is selected from a group comprising S⁺, C, S⁺═O, N⁺H and P═O;X₁ is selected from a group comprising —CH, —CH₂ and —CH₂O; X₂ is C═O;and Z is a platinum containing compound, wherein the platinum forms apart of the ring.

In some embodiments of the various aspects described herein, Q is

wherein X₁ is (CH₂)_(n); X₂ is C═O; Z is a platinum containing compound,wherein the platinum forms a part of the ring; and n is 0, 1, or 2.

In some embodiments of the various aspects disclosed herein, theplatinum is coordinated to a leaving group via a unique O—Ptmonocarboxylato covalent bond and a ═O→Pt coordinate bond. Further, thepresent disclosure also discloses platinum based compounds wherein theplatinum is coordinated to a leaving group via O—Pt monocarboxylato ordicarboxylato covalent bond(s).

In some embodiments of the various aspects disclosed herein, theplatinum moiety is a platinum (II) or platinum (IV) compound. In someembodiments, the platinum (II) compound is selected from the groupcomprising of DACH-platinum, cisplatin, oxaliplatin, carboplatin,paraplatin, sartraplatin, and various combinations thereof. In someembodiments, the platinum containing compound is Pt(II) compound, Pt(IV)compound or halide containing platinum compound. In a preferredembodiment, the platinum compounds are oxaliplatin.

In some embodiments, Z is

wherein R₁ and R₂ are independently halogen, alkyl, amino, alkylamino,dialkylamino, hydroxyl, alkoxy, thiol, thioalkyl, O-acyl, or anycombinations thereof. In some embodiments, R₁ and R₂, together with thePt atom form an optionally substituted cyclyl or heterocyclyl. In someembodiments, Z is

wherein p is 0, 1, 2, or 3. In one embodiment, p is 2.

In some embodiments, Z is

wherein R¹, R² and R³ are independently halogen, alkyl, amino,alkylamino, dialkylamino, hydroxyl, alkoxy, thiol, thioalkyl, O-acyl,-linker-lipid, or any combinations thereof. In some embodiments, R₁ andR₂ together with the Pt atom or R₂ and R₃ together with the Pt atom forman optionally substituted cyclyl or heterocyclyl. In one embodiment, R₁and R₂ together with the Pt atom and R₂ and R₃ together with the Pt atomform an optionally substituted cyclyl or heterocyclyl.

In some embodiments, Z is

R₁ is halogen, alkyl, amino, alkylamino, dialkylamino, hydroxyl, alkoxy,thiol, thioalkyl, O-acyl, or any combinations thereof; and p is 0, 1, 2,or 3. In some further embodiments of this, R₁ is halogen —Cl, —NCS,—O═S(CH₃)₂, —SCH₃, or -linker-lipid. In one embodiment, p is 2.

In some embodiments, Z is

In some embodiments, Z is

wherein R₁, R₂, R₃, R₄ and R₅ are independently halogen, alkyl, amino,alkylamino, dialkylamino, hydroxyl, alkoxy, thiol, thioalkyl, O-acyl,-linker-lipid, or any combinations thereof. In some embodiments, R₁ andR₂ together with the Pt atom form an optionally substituted cyclyl orheterocyclyl. In some embodiments, R₁ and R₂ together with the Pt atomform an optionally substituted cyclyl or heterocyclyl. In oneembodiment, R₁ and R₂ together with the Pt atom form an optionallysubstituted cyclyl or heterocyclyl and R₃ and R₄ together with the Ptatom form an optionally substituted cyclyl or heterocyclyl. In someembodiments, R₅ is OH, OC(O)CH₃, or OC(O)-phenyl.

In some embodiments, Z is

wherein p and q are independently 0, 1, 2, or 3. In some embodiments, pis 2. In some embodiments, q is 2. In one embodiment, p and q are both2.

In one embodiment, Z is

wherein p and q are both 2; and R₅ is OH, OC(O)CH₃, or OC(O)-phenyl.

In some embodiments, the platinum (II) compound comprises at least twonitrogen atoms, where said nitrogen atoms are directly linked toplatinum. In a further embodiment, the two nitrogen atoms are linked toeach other via an optionally substituted linker, e.g. acyclic or cycliclinker. A cyclic linker means a linking moiety that comprises at leastone ring structure. Cyclic linkers can be aryl, heteroaryl, cyclyl orheterocyclyl.

In some embodiments, Q is

wherein R₁ and R₂ are independently halogen, alkyl, amino, alkylamino,dialkylamino, hydroxyl, alkoxy, thiol, thioalkyl, O-acyl, or anycombinations thereof. In some embodiments, R₁ and R₂, together with thePt atom form an optionally substituted cyclyl or heterocyclyl.

In some embodiments, Q is

wherein p is 0, 1, 2, or 3. In one embodiment, p is 2.

In some embodiments, Q is

wherein R¹, R² and R³ are independently halogen, alkyl, amino,alkylamino, dialkylamino, hydroxyl, alkoxy, thiol, thioalkyl, O-acyl, orany combinations thereof. In some embodiments, R₁ and R₂ together withthe Pt atom or R₂ and R₃ together with the Pt atom form an optionallysubstituted cyclyl or heterocyclyl. In one embodiment, R₁ and R₂together with the Pt atom and R₂ and R₃ together with the Pt atom forman optionally substituted cyclyl or heterocyclyl.

In some embodiments, Q is

R₁ is halogen, alkyl, amino, alkylamino, dialkylamino, hydroxyl, alkoxy,thiol, thioalkyl, O-acyl, or any combinations thereof; and p is 0, 1, 2,or 3. In some further embodiments of this, R₁ is halogen —Cl, —NCS,—O═S(CH₃)₂, —SCH₃, or -linker-lipid. In one embodiment, p is 2.

In some embodiments, Q is

In some embodiments, Q is

wherein R₁, R₂, R₃, R₄ and R₅ are independently halogen, alkyl, amino,alkylamino, dialkylamino, hydroxyl, alkoxy, thiol, thioalkyl, O-acyl, orany combinations thereof. In some embodiments, R₁ and R₂ together withthe Pt atom form an optionally substituted cyclyl or heterocyclyl. Insome embodiments, R₁ and R₂ together with the Pt atom form an optionallysubstituted cyclyl or heterocyclyl. In one embodiment, R₁ and R₂together with the Pt atom form an optionally substituted cyclyl orheterocyclyl and R₃ and R₄ together with the Pt atom form an optionallysubstituted cyclyl or heterocyclyl. In some embodiments, R₅ is OH,OC(O)CH₃, or OC(O)-phenyl.

In some embodiments, Q is

wherein p and q are independently 0, 1, 2, or 3. In some embodiments, pis 2. In some embodiments, q is 2. In one embodiment, p and q are both2.

In one embodiment, Q is

wherein p and q are both 2; and R₅ is OH, OC(O)CH₃, or OC(O)-phenyl.

The term “lipid” is used in the conventional sense and includescompounds of varying chain length, from as short as about 2 carbon atomsto as long as about 28 carbon atoms. Additionally, the compounds may besaturated or unsaturated and in the form of straight- or branched-chainsor in the form of unfused or fused ring structures. Exemplary lipidsinclude, but are not limited to, fats, waxes, sterols, steroids, bileacids, fat-soluble vitamins (such as A, D, E, and K), monoglycerides,diglycerides, phospholipids, glycolipids, sulpholipids, aminolipids,chromolipids (lipochromes), glycerophospholipids, sphingolipids,prenollipids, saccharolipids, polyketides, and fatty acids.

Without limitations the lipid can be selected from the group consistingof sterol lipids, fatty acids, fatty alcohols, glycerolipids (e.g.,monoglycerides, diglycerides, and triglycerides), phospholipids,glycerophospholipids, sphingolipids, prenol lipids, saccharolipids,polyketides, and any combination thereof. The lipid can be apolyunsaturated fatty acid or alcohol. The term “polyunsaturated fattyacid” or “polyunsaturated fatty alcohol” as used herein means a fattyacid or alcohol with two or more carbon-carbon double bonds in itshydrocarbon chain. The lipid can also be a highly unsaturated fatty acidor alcohol. The term “highly polyunsaturated fatty acid” or “highlypolyunsaturated fatty alcohol” as used herein means a fatty acid oralcohol having at least 18 carbon atoms and at least 3 double bonds. Thelipid can be an omega-3 fatty acid. The term “omega-3 fatty acid” asused herein means a polyunsaturated fatty acid whose first double bondoccurs at the third carbon-carbon bond from the end opposite the acidgroup.

In some embodiments, the lipid can be selected from the group consistingof 1,3-propanediol dicaprylate/dicaprate; 10-undecenoic acid;1-dotriacontanol; 1-heptacosanol; 1-nonacosanol; 2-ethyl hexanol;androstanes; arachidic acid; arachidonic acid; arachidyl alcohol;behenic acid; behenyl alcohol; Capmul MCM C10; capric acid; capricalcohol; capryl alcohol; caprylic acid; caprylic/capric acid ester ofsaturated fatty alcohol C12-C18; caprylic/capric triglyceride;caprylic/capric triglyceride; ceramide phosphorylcholine (Sphingomyelin,SPH); ceramide phosphorylethanolamine (Sphingomyelin, Cer-PE); ceramidephosphorylglycerol; ceroplastic acid; cerotic acid; cerotic acid; cerylalcohol; cetearyl alcohol; Ceteth-10; cetyl alcohol; cholanes;cholestanes; cholesterol; cis-11-eicosenoic acid; cis-11-octadecenoicacid; cis-13-docosenoic acid; cluytyl alcohol; coenzyme Q10 (CoQ10);dihomo-γ-linolenic; docosahexaenoic acid; egg lecithin; eicosapentaenoicacid; eicosenoic acid; elaidic acid; elaidolinolenyl alcohol;elaidolinoleyl alcohol; elaidyl alcohol; erucic acid; erucyl alcohol;estranes; ethylene glycol distearate (EGDS); geddic acid; geddylalcohol; glycerol distearate (type I) EP (Precirol ATO 5); glyceroltricaprylate/caprate; glycerol tricaprylate/caprate (CAPTEX® 355 EP/NF);glyceryl monocaprylate (Capmul MCM C8 EP); glyceryl triacetate; glyceryltricaprylate; glyceryl tricaprylate/caprate/laurate; glyceryltricaprylate/tricaprate; glyceryl tripalmitate (Tripalmitin);henatriacontylic acid; heneicosyl alcohol; heneicosylic acid;heptacosylic acid; heptadecanoic acid; heptadecyl alcohol;hexatriacontylic acid; isostearic acid; isostearyl alcohol; lacceroicacid; lauric acid; lauryl alcohol; lignoceric acid; lignoceryl alcohol;linoelaidic acid; linoleic acid; linolenyl alcohol; linoleyl alcohol;margaric acid; mead; melissic acid; melissyl alcohol; montanic acid;montanyl alcohol; myricyl alcohol; myristic acid; myristoleic acid;myristyl alcohol; neodecanoic acid; neoheptanoic acid; neononanoic acid;nervonic; nonacosylic acid; nonadecyl alcohol; nonadecylic acid;nonadecylic acid; oleic acid; oleyl alcohol; palmitic acid; palmitoleicacid; palmitoleyl alcohol; pelargonic acid; pelargonic alcohol;pentacosylic acid; pentadecyl alcohol; pentadecylic acid; phosphatidicacid (phosphatidate, PA); phosphatidylcholine (lecithin, PC);phosphatidylethanolamine (cephalin, PE); phosphatidylinositol (PI);phosphatidylinositol bisphosphate (PIP2); phosphatidylinositol phosphate(PIP); phosphatidylinositol triphosphate (PIP3); phosphatidylserine(PS); polyglyceryl-6-distearate; pregnanes; propylene glycol dicaprate;propylene glycol dicaprylocaprate; propylene glycol dicaprylocaprate;psyllic acid; recinoleaic acid; recinoleyl alcohol; sapienic acid; soylecithin; stearic acid; stearidonic; stearyl alcohol; tricosylic acid;tridecyl alcohol; tridecylic acid; triolein; undecyl alcohol;undecylenic acid; undecylic acid; vaccenic acid; α-linolenic acid;γ-linolenic acid; a fatty acid salt of 10-undecenoic acid, adapalene,arachidic acid, arachidonic acid, behenic acid, butyric acid, capricacid, caprylic acid, cerotic acid, cis-11-eicosenoic acid,cis-11-octadecenoic acid, cis-13-docosenoic acid, docosahexaenoic acid,eicosapentaenoic acid, elaidic acid, erucic acid, heneicosylic acid,heptacosylic acid, heptadecanoic acid, isostearic acid, lauric acid,lignoceric acid, linoelaidic acid, linoleic acid, montanic acid,myristic acid, myristoleic acid, neodecanoic acid, neoheptanoic acid,neononanoic acid, nonadecylic acid, oleic acid, palmitic acid,palmitoleic acid, pelargonic acid, pentacosylic acid, pentadecylic acid,recinoleaic acid (e.g. zinc recinoleate), sapienic acid, stearic acid,tricosylic acid, tridecylic acid, undecylenic acid, undecylic acid,vaccenic acid, valeric acid, α-linolenic acid, γ-linolenic acid; and anycombinations thereof.

In some embodiments, the lipid is cholesterol or alpha tocopherol.

As used herein, the term “linker” means an organic moiety that connectstwo parts of a compound. Linkers typically comprise a direct bond or anatom such as oxygen or sulfur, a unit such as NR¹, C(O), C(O)NH, C(O)O,NHC(O)O, OC(O)O, SO, SO₂, SO₂NH or a chain of atoms, such as substitutedor unsubstituted alkyl, substituted or unsubstituted alkenyl,substituted or unsubstituted alkynyl, arylalkyl, arylalkenyl,arylalkynyl, heteroarylalkyl, heteroarylalkenyl, heteroarylalkynyl,heterocyclylalkyl, heterocyclylalkenyl, heterocyclylalkynyl, aryl,heteroaryl, heterocyclyl, cycloalkyl, cycloalkenyl, alkylarylalkyl,alkylarylalkenyl, alkylarylalkynyl, alkenylarylalkyl,alkenylarylalkenyl, alkenylarylalkynyl, alkynylarylalkyl,alkynylarylalkenyl, alkynylarylalkynyl, alkylheteroarylalkyl,alkylheteroarylalkenyl, alkylheteroarylalkynyl, alkenylheteroarylalkyl,alkenylheteroarylalkenyl, alkenylheteroarylalkynyl,alkynylheteroarylalkyl, alkynylheteroarylalkenyl,alkynylheteroarylalkynyl, alkylheterocyclylalkyl,alkylheterocyclylalkenyl, alkylhererocyclylalkynyl,alkenylheterocyclylalkyl, alkenylheterocyclylalkenyl,alkenylheterocyclylalkynyl, alkynylheterocyclylalkyl,alkynylheterocyclylalkenyl, alkynylheterocyclylalkynyl, alkylaryl,alkenylaryl, alkynylaryl, alkylheteroaryl, alkenylheteroaryl,alkynylhereroaryl, where one or more methylenes can be interrupted orterminated by O, S, S(O), SO₂, NR¹, C(O), C(O)NH, C(O)O, NHC(O)O,OC(O)O, SO₂NH, cleavable linking group, substituted or unsubstitutedaryl, substituted or unsubstituted heteroaryl, substituted orunsubstituted heterocyclic; where R¹ is hydrogen, acyl, aliphatic orsubstituted aliphatic.

In some embodiments, the linker is a branched linker. The branchpoint ofthe branched linker may be at least trivalent, but can be a tetravalent,pentavalent or hexavalent atom, or a group presenting such multiplevalencies. In some embodiments, the branchpoint is —N, —N(Q)-C, —O—C,—S—C, —SS—C, —C(O)N(Q)-C, —OC(O)N(Q)-C, —N(Q)C(O)—C, or —N(Q)C(O)O—C;wherein Q is independently for each occurrence H or optionallysubstituted alkyl. In some embodiments, the branchpoint is glycerol orderivative thereof.

A cleavable linking group is one which is sufficiently stable outsidethe cell, but which upon entry into a target cell is cleaved to releasethe two parts the linker is holding together. In a preferred embodiment,the cleavable linking group is cleaved at least 10 times or more,preferably at least 100 times faster in the target cell or under a firstreference condition (which can, e.g., be selected to mimic or representintracellular conditions) than in the blood or serum of a subject, orunder a second reference condition (which can, e.g., be selected tomimic or represent conditions found in the blood or serum).

Cleavable linking groups are susceptible to cleavage agents, e.g., pH,redox potential or the presence of degradative molecules. Generally,cleavage agents are more prevalent or found at higher levels oractivities inside cells than in serum or blood. Examples of suchdegradative agents include: redox agents which are selected forparticular substrates or which have no substrate specificity, including,e.g., oxidative or reductive enzymes or reductive agents such asmercaptans, present in cells, that can degrade a redox cleavable linkinggroup by reduction; esterases; amidases; endosomes or agents that cancreate an acidic environment, e.g., those that result in a pH of five orlower; enzymes that can hydrolyze or degrade an acid cleavable linkinggroup by acting as a general acid, peptidases (which can be substratespecific) and proteases, and phosphatases.

A linker can include a cleavable linking group that is cleavable by aparticular enzyme. The type of cleavable linking group incorporated intoa linker can depend on the cell to be targeted. For example, livertargeting ligands can be linked to the cationic lipids through a linkerthat includes an ester group. Liver cells are rich in esterases, andtherefore the linker will be cleaved more efficiently in liver cellsthan in cell types that are not esterase-rich. Other cell-types rich inesterases include cells of the lung, renal cortex, and testis. Linkersthat contain peptide bonds can be used when targeting cell types rich inpeptidases, such as liver cells and synoviocytes.

In some embodiments, cleavable linking group is cleaved at least 1.25,1.5, 1.75, 2, 3, 4, 5, 10, 25, 50, or 100 times faster in the cell (orunder in vitro conditions selected to mimic intracellular conditions) ascompared to blood or serum (or under in vitro conditions selected tomimic extracellular conditions). In some embodiments, the cleavablelinking group is cleaved by less than 90%, 80%, 70%, 60%, 50%, 40%, 30%,20%, 10%, 5%, or 1% in the blood (or in vitro conditions selected tomimic extracellular conditions) as compared to in the cell (or under invitro conditions selected to mimic intracellular conditions).

Exemplary cleavable linking groups include, but are not limited to,redox cleavable linking groups (e.g., —S—S— and —C(R)₂—S—S—, wherein Ris H or C₁-C₆ alkyl and at least one R is C₁-C₆ alkyl such as CH₃ orCH₂CH₃); phosphate-based cleavable linking groups (e.g., —O—P(O)(OR)—O—,—O—P(S)(OR)—O—, —O—P(S)(SR)—O—, —S—P(O)(OR)—O—, —O—P(O)(OR)—S—,—S—P(O)(OR)—S—, —O—P(S)(ORk)-S—, —S—P(S)(OR)—O—, —O—P(O)(R)—O—,—O—P(S)(R)—O—, —S—P(O)(R)—O—, —S—P(S)(R)—O—, —S—P(O)(R)—S—,—O—P(S)(R)—S—, —O—P(O)(OH)—O—, —O—P(S)(OH)—O—, —O—P(S)(SH)—O—,—S—P(O)(OH)—O—, —O—P(O)(OH)—S—, —S—P(O)(OH)—S—, —O—P(S)(OH)—S—,—S—P(S)(OH)—O—, —O—P(O)(H)—O—, —O—P(S)(H)—O—, —S—P(O)(H)—O—,—S—P(S)(H)—O—, —S—P(O)(H)—S—, and —P(S)(H)—S—, wherein R is optionallysubstituted linear or branched C₁-C₁₀ alkyl); acid cleavable linkinggroups (e.g., hydrazones, esters, and esters of amino acids, —C═NN— and—OC(O)—); ester-based cleavable linking groups (e.g., —C(O)O—);peptide-based cleavable linking groups, (e.g., linking groups that arecleaved by enzymes such as peptidases and proteases in cells, e.g.,—NHCHR^(A)C(O)NHCHR^(B)C(O)—, where R^(A) and R^(B) are the R groups ofthe two adjacent amino acids). A peptide based cleavable linking groupcomprises two or more amino acids. In some embodiments, thepeptide-based cleavage linkage comprises the amino acid sequence that isthe substrate for a peptidase or a protease found in cells.

In some embodiments, an acid cleavable linking group is cleavable in anacidic environment with a pH of about 6.5 or lower (e.g., about 6.5,6.0, 5.5, 5.0, or lower), or by agents such as enzymes that can act as ageneral acid.

Linkers according to the present invention include moieties comprisingtwo or more carbon molecules such as, for example, ethylenediamine,ethyleneglycol, glycine, beta-alanine and polyethylene glycol (PEG) ofmolecular weight about 44 to about 200 kD. Further, it is to beunderstood from the present disclosure that the platinum moiety and/orthe lipid may be modified to comprise functional groups for linking tothe linker molecule.

In some embodiments of the various aspects disclosed herein, the linkeris —X—CH₂—X₂—X₁—, wherein X is NH; X₁ is C(O)O, C(O)NH, O(CH₂)—O, NH, orO; X₂ is (CH₂)_(n) or C(O); and n is 0, 1, 2, 3, 4, or 5.

In some other embodiments, the linker is —(CH₂)_(n)O—,—(CH₂)_(n)NHC(O)O—, —(CH₂)_(n)OC(O)NH—, —(CH₂)_(n)C(O)NH(CH₂)_(m)O—,—(CH₂)_(n)O(CH₂)_(m)O—, —(CH₂)_(n)O(O)—, —(CH₂)_(n)NHC(O)(CH₂)_(m)O—, or—(CH₂)_(n)C(O)O—; and n and m are independently 0, 1, 2, 3, 4, or 5.

In still some other embodiments, the linker is —X₃—X₄X₅—X₆—, wherein X₃is CH, CH₂, or O; and X₄, X₅ and X₆ are independently same or differentand are —CH₂O— or O.

In yet some other embodiments, the linker is —CH₂O—.

In some embodiments, the linker is selected from the group consisting ofa bond, —O—, NHCH₂CH₂NHC(O)—, —NHCH₂CH₂NHC(O)O—, —NHCH₂CH₂—,—NHCH₂CH₂O—, —NHCH₂C(O)—, —NHCH₂C(O)O—, —NHCH₂C(O)OCH₂CH₂CH₂—,—NHCH₂C(O)OCH₂CH₂CH₂O—, —NHCH₂C(O)NH—, —CH₂CH₂—, —CH₂CH₂O—,—CH₂CH₂NHC(O)—, —CH₂CH₂NHC(O)O—, —CH₂CH₂O—, —CH₂C(O)NHCH₂CH₂—,—CH₂C(O)NHCH₂CH₂O—, —CH₂CH₂OCH₂CH₂—, —CH₂CH₂OCH₂CH₂O—, —CH₂C(O)—,—CH₂C(O)O—, —CH₂CH₂CH₂—, —CH₂CH₂CH₂O—, ═CH—CH═CH₂—, ═CH—CH═CHCH₂O—,—CH═CHCH₂—, —CH═CHCH₂O—, —OCH₂CH₂O—, —CH₂—, —CH₂O—, —NHC(O)CH₂—,—NHC(O)CH₂O—, —C(O)CH₂—, —C(O)CH₂O—, —OC(O)CH₂—, —OC(O)CH₂O—,—C(O)CH₂CH₂C(O)NHCH₂CH₂—, —OC(O)CH₂CH₂C(O)NHCH₂CH₂—,—C(O)CH₂CH₂C(O)NHCH₂CH₂O—, —OC(O)CH₂CH₂C(O)NHCH₂CH₂O—,—C(O)CH₂CH₂C(O)NHCH₂CH₂NHC(O)—, —OC(O)CH₂CH₂C(O)NHCH₂CH₂NHC(O)—,—C(O)CH₂CH₂C(O)NHCH₂CH₂NHC(O)O—, —OC(O)CH₂CH₂C(O)NHCH₂CH₂NHC(O)O—, andany combinations thereof.

In some embodiments, the platinum based compounds disclosed herein arerepresented by Formula (I):

-   -   wherein,    -   X is NH;    -   X₁ is selected from a group comprising COOH, CONH₂,        O—(CH₂)_(n)—OH, NH₂ and OH;    -   X₂ is (CH₂)_(n) or CO;    -   X₃ is selected from a group comprising (CH₂)_(n), CH₂—NH and        C₄H₈;    -   X₄ is CO or —CH—CH₃;    -   Z is a platinum containing compound, wherein the platinum forms        a part of Formula I ring; and    -   n is 0, 1, or 2.

In some other embodiments, the platinum based compounds disclosed hereinare represented by Formula (II):

-   -   wherein,    -   X is NH or N—CH₂COO⁻;    -   X₁ is selected from a group comprising —(CH₂)_(n)OH,        —(CH₂)_(n)NHCOOH, —(CH₂)_(n)CONH(CH₂)_(n)OH,        (CH₂)_(n)O(CH₂)_(n)OH, (CH₂)_(n)C═O, —(CH₂)_(n)NHCO(CH₂)_(n)OH        and (CH₂)_(n)—COOH;    -   Z is platinum containing compound, wherein the platinum forms a        part of Formula II ring; and    -   n is 0, 1, or 2.

In some embodiments, the platinum based compounds disclosed herein arerepresented by Formula (III):

-   -   wherein,    -   X is selected from a group comprising S⁺, C, S⁺═O, N⁺H and P═O;    -   X₁ is selected from a group comprising —CH, —CH₂ and —CH₂O;    -   X₂ is C═O;    -   X₃ is selected from CH, CH₂ or O;    -   X₄, X₅, X₆ is selected from —CH₂O or O;    -   Z is platinum containing compound, wherein the platinum forms a        part of Formula III ring.

In some embodiments, the platinum based compounds disclosed herein arerepresented by Formula (IV):

-   -   wherein,    -   X is CH₂OH;    -   X₁ is (CH₂)_(n);    -   X₂ is C═O;    -   Z is platinum containing compound, wherein the platinum forms a        part of Formula IV ring; and    -   n is 0, 1, or 2.

Exemplary compounds of Formula (I) include, but are not limited to, thefollowing compounds:

Exemplary compounds of Formula (II) include, but are not limited to, thefollowing compounds:

Exemplary compounds of Formula (III) include, but are not limited to,the following compounds:

Exemplary compounds of Formula (IV) include, but are not limited to, thefollowing compounds:

The disclosure also provides the following compounds:

In some embodiments, the platinum moiety of the platinum based complexesof the present disclosure is a platinum (IV) compound. Said complexeshaving platinum (IV) compounds are represented as follows.

In some embodiments of the various aspects disclosed herein, thedisclosure provides a platinum (II) compound of Formula (V):

wherein

-   -   X₁, X₂, X₃ and X₄ are selected independently from the group        consisting of O, P, S, Se, Cl, N, C, O-A, O—B, DACH, halides and        chelated or non-chelated dicarboxylato linkage group, and any        combinations thereof;    -   wherein A and B are selected independently from the group        consisting of C, P, S, N, and any combinations thereof; and    -   wherein X₄ is optional.

In some embodiments of the various aspects disclosed herein, theplatinum (II) compound of Formula (V) is selected from a groupcomprising Compounds 43-65 and 73-85 and Compound 95. In a preferredembodiment, the Pt(II) compound is DACH-Pt.

In the above compounds, R¹ is a -linker-lipid and n is 1 unless definedotherwise.

In still another embodiment, a Platinum (II) compound [Compound 95] isalso provided by the present disclosure.

Synthesis of some exemplary compounds of Formula (V) is described in theExamples section.

Without wishing to be bound by a theory, compounds disclosed herein havehigher uptake of platinum in cancer cells relative to cisplatin andoxaliplatin. As shown in FIG. 8, the uptake of cisplatin and oxaliplatinare similar in MDA-MB-231 cells. However, of the IO-compounds testedshowed higher uptake (˜7-20 fold). These results indicate that whenadministered at platinum equivalent concentrations, the uptake ofIO-compounds is significantly higher in comparison to cisplatin oroxaliplatin in cancer cells. In some embodiments, the compoundsdisclosed herein have about 25%, about 50%, about 75%, about 1-fold,about 5-folds, about 10-folds, about 15-folds, about 20-folds, about25-folds or higher platinum uptake in cancer cells relative to cisplatinor oxaliplatin at equivalent dosage.

In addition, the compounds disclosed herein also have higheraccumulation of platinum in tissue, such as, but not limited to a tumor,relative to cisplatin and oxaliplatin when dosed at equivalent amount.For example, the compounds disclosed herein have about 25%, about 50%,about 75%, about 1-fold, about 5-folds, about 10-folds, about 15-folds,about 20-folds, about 25-folds or higher platinum accumulation tissuerelative to cisplatin or oxaliplatin when dosed at equivalent amounts.

The present disclosure relates to the synthesis of a series of platinumbased nanoparticles wherein, the diaminocyclohexyl-Pt (DACH-Pt) has amonocarboxylated covalent bond through a carboxylic acid and aco-ordination bond with amide oxygen. Dicarbonyl molecules (dicarboxylicacids) such as succinic acid, malonic acid and oxalic acid are usedwhich eventually form seven, six and five member rings with platinum(II) respectively. The linker between the platinum ring and cholesterolhelps in forming linkages selected from a group comprising carbamatelinkage (compounds 1, 2, 3), ether linkage (compounds 6, 4, 5) or thelikes or any combinations thereof. Therefore, some of the embodiments ofthe present disclosure relates to compounds represented by the generalbackbone: lipid-linker-dicarbonyl. These molecules are used to complexplatinum compounds such as DACH-Pt, oxaliplatin, cisplatin, platinumcontaining carbenes or other platinates and platinum compounds, throughcovalent and/or coordination bonds.

In an embodiment of the present disclosure, several variants of platinumbased compounds such as racemates, diastereomers and the likes are alsoprovided (for example, Compounds 1-6).

In an embodiment of the present disclosure, any molecule that has twocarbonyl groups may be used. In one embodiment, the dicarbonyl moleculeis a dicarboxylic acid, such as, for example, succinic acid, malonicacid or oxalic acid.

The disclosure also provides particles comprising one or more of theplatinum based compounds described herein. Generally, the particledisclosed herein can be of any shape or form, e.g., spherical, rod,elliptical, cylindrical, capsule, or disc; and these particles can bepart of a network or an aggregate.

In some embodiments, the particle is a microparticle or a nanoparticle.As used herein, the term “microparticle” refers to a particle having aparticle size of about 1 μm to about 1000 μm. As used herein, the term“nanoparticle” refers to particle having a particle size of about 0.1 nmto about 1000 nm. Generally, the particles have any size from nm tomillimeters. In some embodiments, the particles can have a size rangingfrom about 5 nm to about 5000 nm. In some embodiments, the particleshave an average diameter of from about 50 nm to about 2500 nm. In someembodiments, the particles have an average diameter of from about 100 nmto about 2000 nm. In some embodiments, the particles have an averagediameter of from about 150 nm to about 1700 nm. In some embodiments, theparticles have an average diameter of from about 200 nm to about 1500nm. In some embodiment, the particles have an average diameter of about260 nm. In one embodiment, the particles have an average diameter ofabout 30 nm to about 150 nm. In some embodiments, the particle have anaverage diameter of about 100 nm to about 1000 nm, from about 200 nm toabout 800 nm, from about 200 nm to about 700 nm, or from about 300 nm toabout 700 nm.

In some embodiments, the particle has an average size of about 50 toabout 1000 nm. In a further embodiment, the nanoparticles of the presentinvention are in the range of about 50 to about 500 nm. In anotherembodiment, the nanoparticles of the present invention are in the rangeof about 50 to about 500 nm (FIG. 5). In one embodiment, the particlehas a size of about 500 nm.

It will be understood by one of ordinary skill in the art that particlesusually exhibit a distribution of particle sizes around the indicated“size.” Unless otherwise stated, the term “particle size” as used hereinrefers to the mode of a size distribution of particles, i.e., the valuethat occurs most frequently in the size distribution. Methods formeasuring the particle size are known to a skilled artisan, e.g., bydynamic light scattering (such as photocorrelation spectroscopy, laserdiffraction, low-angle laser light scattering (LALLS), and medium-anglelaser light scattering (MALLS)), light obscuration methods (such asCoulter analysis method), or other techniques (such as rheology, andlight or electron microscopy).

In some embodiments, the particles can be substantially spherical. Whatis meant by “substantially spherical” is that the ratio of the lengthsof the longest to the shortest perpendicular axes of the particle crosssection is less than or equal to about 1.5. Substantially spherical doesnot require a line of symmetry. Further, the particles can have surfacetexturing, such as lines or indentations or protuberances that are smallin scale when compared to the overall size of the particle and still besubstantially spherical. In some embodiments, the ratio of lengthsbetween the longest and shortest axes of the particle is less than orequal to about 1.5, less than or equal to about 1.45, less than or equalto about 1.4, less than or equal to about 1.35, less than or equal toabout 1.30, less than or equal to about 1.25, less than or equal toabout 1.20, less than or equal to about 1.15 less than or equal to about1.1. Without wishing to be bound by a theory, surface contact isminimized in particles that are substantially spherical, which minimizesthe undesirable agglomeration of the particles upon storage. Manycrystals or flakes have flat surfaces that can allow large surfacecontact areas where agglomeration can occur by ionic or non-ionicinteractions. A sphere permits contact over a much smaller area.

In some embodiments, the particles have substantially the same particlesize. Particles having a broad size distribution where there are bothrelatively big and small particles allow for the smaller particles tofill in the gaps between the larger particles, thereby creating newcontact surfaces. A broad size distribution can result in larger spheresby creating many contact opportunities for binding agglomeration. Theparticles described herein are within a narrow size distribution,thereby minimizing opportunities for contact agglomeration. What ismeant by a “narrow size distribution” is a particle size distributionthat has a ratio of the volume diameter of the 90th percentile of thesmall spherical particles to the volume diameter of the 10th percentileless than or equal to 5. In some embodiments, the volume diameter of the90th percentile of the small spherical particles to the volume diameterof the 10th percentile is less than or equal to 4.5, less than or equalto 4, less than or equal to 3.5, less than or equal to 3, less than orequal to 2.5, less than or equal to 2, less than or equal to 1.5, lessthan or equal to 1.45, less than or equal to 1.40, less than or equal to1.35, less than or equal to 1.3, less than or equal to 1.25, less thanor equal to 1.20, less than or equal to 1.15, or less than or equal to1.1.

Geometric Standard Deviation (GSD) can also be used to indicate thenarrow size distribution. GSD calculations involved determining theeffective cutoff diameter (ECD) at the cumulative less than percentagesof 15.9% and 84.1%. GSD is equal to the square root of the ratio of theECD less than 84.17% to ECD less than 15.9%. The GSD has a narrow sizedistribution when GSD<2.5. In some embodiments, GSD is less than 2, lessthan 1.75, or less than 1.5. In one embodiment, GSD is less than 1.8.

In addition to the platinum compounds disclosed herein, the particle cancomprise co-lipids and/stabilizers. Additional lipids can be included inthe particles for a variety of purposes, such as to prevent lipidoxidation, to stabilize the bilayer, to reduce aggregation duringformation or to attach ligands onto the particle surface. Any of anumber of additional lipids and/or other components can be present,including amphipathic, neutral, cationic, anionic lipids, andprogrammable fusion lipids. Such lipids and/or components can be usedalone or in combination. One or more components of particle can comprisea ligand, e.g., a targeting ligand.

In some embodiments, the particle further comprises a phospholipid.Without limitations, the phospholipids can be of natural origin, such asegg yolk or soybean phospholipids, or synthetic or semisynthetic origin.The phospholipids can be partially purified or fractionated to comprisepure fractions or mixtures of phosphatidyl cholines, phosphatidylcholines with defined acyl groups having 6 to 22 carbon atoms,phosphatidyl ethanolamines, phosphatidyl inositols, phosphatidic acids,phosphatidyl serines, sphingomyelin or phosphatidyl glycerols. Suitablephospholipids include, but are not limited to, phosphatidylcholine,phosphatidylglycerol, lecithin, β,γ-dipalmitoyl-α-lecithin,sphingomyelin, phosphatidylserine, phosphatidic acid,N-(2,3-di(9-(Z)-octadecenyloxy))-prop-1-yl-N,N,N-trimethylammoniumchloride, phosphatidylethanolamine, lysolecithin,lysophosphatidylethanolamine, phosphatidylinositol, cephalin,cardiolipin, cerebrosides, dicetylphosphate,dioleoylphosphatidylcholine, dipalmitoylphosphatidylcholine,dipalmitoylphosphatidylglycerol, dioleoylphosphatidylglycerol,palmitoyl-oleoyl-phosphatidylcholine, di-stearoyl-phosphatidylcholine,stearoyl-palmitoyl-phosphatidylcholine,di-palmitoyl-phosphatidylethanolamine,di-stearoyl-phosphatidylethanolamine, di-myrstoyl-phosphatidylserine,di-oleyl-phosphatidylcholine, dimyristoyl phosphatidyl choline (DMPC),dioleoylphosphatidylethanolamine (DOPE),palmitoyloleoylphosphatidylcholine (POPC), egg phosphatidylcholine(EPC), distearoylphosphatidylcholine (DSPC), dioleoylphosphatidylcholine(DOPC), dipalmitoylphosphatidylcholine (DPPC),dioleoylphosphatidylglycerol (DOPG), dipalmitoylphosphatidylglycerol(DPPG), -phosphatidylethanolamine (POPE),dioleoyl-phosphatidylethanolamine4-(N-maleimidomethyl)-cyclohexane-1-carboxylate (DOPE-mal),1-stearoyl-2-oleoyl phosphatidylcholine (SOPC),1,2-distearoyl-sn-glycem-3-phosphoethanolamine (DSPE), and anycombinations thereof. Non-phosphorus containing lipids can also be used.These include, e.g., stearylamine, docecylamine, acetyl palmitate, fattyacid amides, and the like. Other phosphorus-lacking compounds, such assphingolipids, glycosphingolipid families, diacylglycerols, andβ-acyloxyacids, can also be used

In some embodiments, the phospholipid in the particle is selected fromthe group consisting of 1,2-didecanoyl-sn-glycero-3-phosphocholine;1,2-dierucoyl-sn-glycero-3-phosphate (sodium salt);1,2-dierucoyl-sn-glycero-3-phosphocholine;1,2-dierucoyl-sn-glycero-3-phosphoethanolamine;1,2-dierucoyl-sn-glycero-3[phospho-rac-(1-glycerol) (sodium salt);1,2-dilinoleoyl-sn-glycero-3-phosphocholine;1,2-dilauroyl-sn-glycero-3-phosphate (sodium salt);1,2-dilauroyl-sn-glycero-3-phosphocholine;1,2-dilauroyl-sn-glycero-3-phosphoethanolamine;1,2-dilauroyl-sn-glycero-3[phospho-rac-(1-glycerol) (sodium salt);1,2-dilauroyl-sn-glycero-3[phospho-rac-(1-glycerol) (ammonium salt);1,2-dilauroyl-sn-glycero-3-phosphoserine (sodium salt);1,2-dimyristoyl-sn-glycero-3-phosphate (sodium salt);1,2-dimyristoyl-sn-glycero-3-phosphocholine;1,2-dimyristoyl-sn-glycero-3-phosphoethanolamine;1,2-dimyristoyl-sn-glycero-3[phospho-rac-(1-glycerol) (sodium salt);1,2-dimyristoyl-sn-glycero-3[phospho-rac-(1-glycerol) (ammonium salt);1,2-dimyristoyl-sn-glycero-3[phospho-rac-(1-glycerol) (sodium/ammoniumsalt); 1,2-dimyristoyl-sn-glycero-3-phosphoserine (sodium salt);1,2-dioleoyl-sn-glycero-3-phosphate (sodium salt);1,2-dioleoyl-sn-glycero-3-phosphocholine;1,2-dioleoyl-sn-glycero-3-phosphoethanolamine;1,2-dioleoyl-sn-glycero-3[phospho-rac-(1-glycerol) (sodium salt);1,2-dioleoyl-sn-glycero-3-phosphoserine (sodium salt);1,2-dipalmitoyl-sn-glycero-3-phosphate (sodium salt);1,2-dipalmitoyl-sn-glycero-3-phosphocholine;1,2-dipalmitoyl-sn-glycero-3-phosphoethanolamine;1,2-dipalmitoyl-sn-glycero-3[phospho-rac-(1-glycerol) (sodium salt);1,2-dipalmitoyl-sn-glycero-3[phospho-rac-(1-glycerol) (ammonium salt);1,2-dipalmitoyl-sn-glycero-3-phosphoserine (sodium salt);1,2-distearoyl-sn-glycero-3-phosphate (sodium salt);1,2-distearoyl-sn-glycero-3-phosphocholine;1,2-distearoyl-sn-glycero-3-phosphoethanolamine;1,2-distearoyl-sn-glycero-3[phospho-rac-(1-glycerol) (sodium salt);1,2-distearoyl-sn-glycero-3[phospho-rac-(1-glycerol) (ammonium salt);1,2-distearoyl-sn-glycero-3-phosphoserine (sodium salt); Egg-PC;Hydrogenated Egg PC; hydrogenated soy PC;1-myristoyl-sn-glycero-3-phosphocholine;1-palmitoyl-sn-glycero-3-phosphocholine;1-stearoyl-sn-glycero-3-phosphocholine;1-myristoyl-2-palmitoyl-sn-glycero 3-phosphocholine;1-myristoyl-2-stearoyl-sn-glycero-3-phosphocholine;1-palmitoyl-2-myristoyl-sn-glycero-3-phosphocholine;1-palmitoyl-2-oleoyl-sn-glycero-3-phosphocholine;1-palmitoyl-2-oleoyl-sn-glycero-3-phosphoethanolamine;1-palmitoyl-2-oleoyl-sn-glycero-3[phospho-rac-(1-glycerol)](sodiumsalt); 1-palmitoyl-2-stearoyl-sn-glycero-3-phosphocholine;1-stearoyl-2-myristoyl-sn-glycero-3-phosphocholine;1-stearoyl-2-oleoyl-sn-glycero-3-phosphocholine; and1-stearoyl-2-palmitoyl-sn-glycero-3-phosphocholine. In some embodiments,the phospholipid is SPOC, egg PC, or Hydrogenated Soy PC (HSPC). In one,the phospholipid in the composition is HSPC.

In some embodiments, the particle further comprises a polyethyleneglycol (PEG). The PEG can be included in the particle by itself orconjugated with a component present in the particle. For example, thePEG can be conjugated with the platinum based compound or aco-lipid/stabilizer component of the particle. In some embodiments, thePEG is conjugated with a co-lipid component of the particle. Withoutlimitations, the PEG can be conjugated with any co-lipid. For example,the PEG conjugated co-lipid can be selected from the group consisting ofPEG conjugated diacylglycerols and dialkylglycerols, PEG-conjugatedphosphatidylethanolamine, PEG conjugated to phosphatidic acid, PEGconjugated ceramides (see, U.S. Pat. No. 5,885,613), PEG conjugateddialkylamines, PEG conjugated 1,2-diacyloxypropan-3-amines, and PEGconjugated to 1,2-distearoyl-sn-glycem-3-phosphoethanolamine (DSPE), andany combinations thereof. In some embodiments, the PEG conjugated lipidis 1,2-distearoyl-sn-glycem-3-phosphoethanolamine-N-[amino(polyethyleneglycol)-2000] (DSPE-PEG2000).

In some embodiments, the particle further comprises a surfactant.Surfactants find wide application in formulations such as emulsions(including microemulsions) and liposomes. The most common way ofclassifying and ranking the properties of the many different types ofsurfactants, both natural and synthetic, is by the use of thehydrophile/lipophile balance (HLB). The nature of the hydrophilic group(also known as the “head”) provides the most useful means forcategorizing the different surfactants used in formulations (Rieger, inPharmaceutical Dosage Forms, Marcel Dekker, Inc., New York, N.Y., 1988,p. 285).

If the surfactant molecule is not ionized, it is classified as anonionic surfactant. Nonionic surfactants find wide application inpharmaceutical and cosmetic products and are usable over a wide range ofpH values. In general their HLB values range from 2 to about 18depending on their structure. Nonionic surfactants include nonionicesters such as ethylene glycol esters, propylene glycol esters, glycerylesters, polyglyceryl esters, sorbitan esters, sucrose esters, andethoxylated esters. Nonionic alkanolamides and ethers such as fattyalcohol ethoxylates, propoxylated alcohols, and ethoxylated/propoxylatedblock polymers are also included in this class. The polyoxyethylenesurfactants are the most popular members of the nonionic surfactantclass.

If the surfactant molecule carries a negative charge when it isdissolved or dispersed in water, the surfactant is classified asanionic. Anionic surfactants include carboxylates such as soaps, acyllactylates, acyl amides of amino acids, esters of sulfuric acid such asalkyl sulfates and ethoxylated alkyl sulfates, sulfonates such as alkylbenzene sulfonates, acyl isethionates, acyl taurates andsulfosuccinates, and phosphates. The most important members of theanionic surfactant class are the alkyl sulfates and the soaps.

If the surfactant molecule carries a positive charge when it isdissolved or dispersed in water, the surfactant is classified ascationic. Cationic surfactants include quaternary ammonium salts andethoxylated amines. The quaternary ammonium salts are the most usedmembers of this class.

If the surfactant molecule has the ability to carry either a positive ornegative charge, the surfactant is classified as amphoteric. Amphotericsurfactants include acrylic acid derivatives, substituted alkylamides,N-alkylbetaines and phosphatides.

The use of surfactants in drug products, formulations and in emulsionshas been reviewed (Rieger, in Pharmaceutical Dosage Forms, MarcelDekker, Inc., New York, N.Y., 1988, p. 285).

In some embodiments, the particle can further comprise a cationic lipid.Exemplary cationic lipids include, but are not limited to,N,N-dioleyl-N,N-dimethylammonium chloride (DODAC),N,N-distearyl-N,N-dimethylammonium bromide (DDAB),N-(1-(2,3-dioleoyloxy)propyl)-N,N,N-trimethylammonium chloride (DOTAP),N-(1-(2,3-dioleyloxy)propyl)-N,N,N-trimethylammonium chloride (DOTMA),N,N-dimethyl-2,3-dioleyloxy)propylamine (DODMA),1,2-diLinoleyloxy-N,N-dimethylaminopropane (DLinDMA),1,2-dilinolenyloxy-N,N-dimethylaminopropane (DLenDMA),1,2-dilinoleylcarbamoyloxy-3-dimethylaminopropane (DLin-C-DAP),1,2-dilinoleyoxy-3-(dimethylamino)acetoxypropane (DLin-DAC),1,2-dilinoleyoxy-3-morpholinopropane (DLin-MA),1,2-dilinoleoyl-3-dimethylaminopropane (DLinDAP),1,2-dilinoleylthio-3-dimethylaminopropane (DLin-S-DMA),1-linoleoyl-2-linoleyloxy-3-dimethylaminopropane (DLin-2-DMAP),1,2-dilinoleyloxy-3-trimethylaminopropane chloride salt (DLin-TMA.C1),1,2-dilinoleoyl-3-trimethylaminopropane chloride salt (DLin-TAP.C1),1,2-Dilinoleyloxy-3-(N-methylpiperazino)propane (DLin-MPZ), or3-(N,N-dilinoleylamino)-1,2-propanediol (DLinAP),3-(N,N-dioleylamino)-1,2-propanedio (DOAP),1,2-dilinoleyloxo-3-(2-N,N-dimethylamino)ethoxypropane (DLin-EG-DMA),1,2-dilinolenyloxy-N,N-dimethylaminopropane (DLinDMA),2,2-dilinoleyl-4-dimethylaminomethyl-[1,3]-dioxolane (DLin-K-DMA) oranalogs thereof,(3aR,5s,6aS)-N,N-dimethyl-2,2-di((9Z,12Z)-octadeca-9,12-dienyl)tetrahydro-3aH-cyclopenta[d][1,3]dioxol-5-amine(ALN100), (6Z,9Z,28Z,31Z)-heptatriaconta-6,9,28,31-tetraen-19-yl4-(dimethylamino)butanoate (MC3),1,1′-(2-(4-(2-((2-(bis(2-hydroxydodecyl)amino)ethyl)(2-hydroxydodecyl)amino)ethyl)piperazin-1-yl)ethylazanediyl)didodecan-2-ol(Tech Gi), or a mixture thereof.

In some embodiments, the particle further comprises a non-cationiclipid. The non-cationic lipid can be an anionic lipid or a neutral lipidincluding, but not limited to, distearoylphosphatidylcholine (DSPC),dioleoylphosphatidylcholine (DOPC), dipalmitoylphosphatidylcholine(DPPC), dioleoylphosphatidylglycerol (DOPG),dipalmitoylphosphatidylglycerol (DPPG),dioleoyl-phosphatidylethanolamine (DOPE),palmitoyloleoylphosphatidylcholine (POPC),palmitoyloleoylphosphatidylethanolamine (POPE),dioleoyl-phosphatidylethanolamine4-(N-maleimidomethyl)-cyclohexane-1-carboxylate (DOPE-mal), dipalmitoylphosphatidyl ethanolamine (DPPE), dimyristoylphosphoethanolamine (DMPE),distearoyl-phosphatidyl-ethanolamine (DSPE), 16-O-monomethyl PE,16-O-dimethyl PE, 18-1-trans PE,1-stearoyl-2-oleoyl-phosphatidyethanolamine (SOPE), cholesterol, or amixture thereof.

The conjugated lipids that inhibits aggregation of particles can also beincluded in the particles disclosed herein. Such lipids include, but arenot limited to, a polyethyleneglycol (PEG)-lipid including, withoutlimitation, a PEG-diacylglycerol (DAG), a PEG-dialkyloxypropyl (DAA), aPEG-phospholipid, a PEG-ceramide (Cer), or a mixture thereof. ThePEG-DAA conjugate can be, for example, a PEG-dilauryloxypropyl (C₁₂), aPEG-dimyristyloxypropyl (C₁₄), a PEG-dipalmityloxypropyl (C₁₆), or aPEG-distearyloxypropyl (Cis). The conjugated lipid that preventsaggregation of particles can be from 0.01 mol % to about 20 mol % orabout 2 mol % of the total lipid present in the particle.

In some embodiments, the particle is in the form of a liposome, vesicle,or emulsion. As used herein, the term “liposome” encompasses anycompartment enclosed by a lipid layer. Liposomes can have one or morelipid membranes. Liposomes can be characterized by membrane type and bysize. Small unilamellar vesicles (SUVs) have a single membrane andtypically range between 0.02 and 0.05 μm in diameter; large unilamellarvesicles (LUVS) are typically larger than 0.05 μm. Oligolamellar largevesicles and multilamellar vesicles have multiple, usually concentric,membrane layers and are typically larger than 0.1 μm. Liposomes withseveral nonconcentric membranes, i.e., several smaller vesiclescontained within a larger vesicle, are termed multivesicular vesicles.

In order to form a liposome the lipid molecules comprise elongatednon-polar (hydrophobic) portions and polar (hydrophilic) portions. Thehydrophobic and hydrophilic portions of the molecule are preferablypositioned at two ends of an elongated molecular structure. When suchlipids are dispersed in water they spontaneously form bilayer membranesreferred to as lamellae. The lamellae are composed of two mono layersheets of lipid molecules with their non-polar (hydrophobic) surfacesfacing each other and their polar (hydrophilic) surfaces facing theaqueous medium. The membranes formed by the lipids enclose a portion ofthe aqueous phase in a manner similar to that of a cell membraneenclosing the contents of a cell. Thus, the bilayer of a liposome hassimilarities to a cell membrane without the protein components presentin a cell membrane.

A liposome composition can be prepared by a variety of methods that areknown in the art. See e.g., U.S. Pat. Nos. 4,235,871, 4,897,355 and5,171,678; published PCT applications WO 96/14057 and WO 96/37194;Felgner, P. L. et al., Proc. Nat. Acad. Sci., USA (1987) 8:7413-7417,Bangham, et al. M Mol. Biol. (1965) 23:238, Olson, et al. Biochim.Biophys. Acta (1979) 557:9, Szoka, et al. Proc. Nat. Acad. Sci. (1978)75: 4194, Mayhew, et al. Biochim. Biophys. Acta (1984) 775:169, Kim, etal. Biochim. Biophys. Acta (1983) 728:339, and Fukunaga, et al.Endocrinol. (1984) 115:757, content of all of which is incorporatedherein by reference in its entirety.

The liposomes can be prepared to have substantially homogeneous sizes ina selected size range. One effective sizing method involves extruding anaqueous suspension of the liposomes through a series of polycarbonatemembranes having a selected uniform pore size; the pore size of themembrane will correspond roughly with the largest sizes of liposomesproduced by extrusion through that membrane. See e.g., U.S. Pat. No.4,737,323, content of which is incorporated herein by reference in itsentirety.

The particles can also be in the form of an emulsion. Emulsions aretypically heterogeneous systems of one liquid dispersed in another inthe form of droplets (Idson, in Pharmaceutical Dosage Forms, Lieberman,Rieger and Banker (Eds.), 1988, Marcel Dekker, Inc., New York, N.Y.,volume 1, p. 199; Rosoff, in Pharmaceutical Dosage Forms, Lieberman,Rieger and Banker (Eds.), 1988, Marcel Dekker, Inc., New York, N.Y.,Volume 1, p. 245; Block in Pharmaceutical Dosage Forms, Lieberman,Rieger and Banker (Eds.), 1988, Marcel Dekker, Inc., New York, N.Y.,volume 2, p. 335; Higuchi et al., in Remington's PharmaceuticalSciences, Mack Publishing Co., Easton, Pa., 1985, p. 301). Emulsions areoften biphasic systems comprising two immiscible liquid phasesintimately mixed and dispersed with each other. In general, emulsionsmay be of either the water-in-oil (w/o) or the oil-in-water (o/w)variety. When an aqueous phase is finely divided into and dispersed asminute droplets into a bulk oily phase, the resulting composition iscalled water-in-oil (w/o) emulsion. Alternatively, when an oily phase isfinely divided into and dispersed as minute droplets into a bulk aqueousphase, the resulting composition is called an oil-in-water (o/w)emulsion. Emulsions can contain additional components in addition to thedispersed phases, and the conjugate disclosed herein can be present as asolution in either the aqueous phase or the oily phase or itself as aseparate phase. Pharmaceutical excipients such as emulsifiers,stabilizers, dyes, and anti-oxidants can also be present in emulsions asneeded. Pharmaceutical emulsions can also be multiple emulsions that arecomprised of more than two phases such as, for example, in the case ofoil-in-water-in-oil (o/w/o) and water-in-oil-in-water (w/o/w) emulsions.Such complex formulations often provide certain advantages that simplebinary emulsions do not. Multiple emulsions in which individual oildroplets of an o/w emulsion enclose small water droplets constitute aw/o/w emulsion. Likewise a system of oil droplets enclosed in globulesof water stabilized in an oily continuous phase provides an o/w/oemulsion.

Emulsions are characterized by little or no thermodynamic stability.Often, the dispersed or discontinuous phase of the emulsion is welldispersed into the external or continuous phase and maintained in thisform through the means of emulsifiers or the viscosity of theformulation. Either of the phases of the emulsion may be a semisolid ora solid, as is the case of emulsion-style ointment bases and creams.Other means of stabilizing emulsions entail the use of emulsifiers thatmay be incorporated into either phase of the emulsion. Emulsifiers canbroadly be classified into four categories: synthetic surfactants,naturally occurring emulsifiers, absorption bases, and finely dispersedsolids (Idson, in Pharmaceutical Dosage Forms, Lieberman, Rieger andBanker (Eds.), 1988, Marcel Dekker, Inc., New York, N.Y., volume 1, p.199).

Synthetic surfactants, also known as surface active agents, have foundwide applicability in the formulation of emulsions and have beenreviewed in the literature (Rieger, in Pharmaceutical Dosage Forms,Lieberman, Rieger and Banker (Eds.), 1988, Marcel Dekker, Inc., NewYork, N.Y., volume 1, p. 285; Idson, in Pharmaceutical Dosage Forms,Lieberman, Rieger and Banker (Eds.), Marcel Dekker, Inc., New York,N.Y., 1988, volume 1, p. 199). Surfactants are typically amphiphilic andcomprise a hydrophilic and a hydrophobic portion. The ratio of thehydrophilic to the hydrophobic nature of the surfactant has been termedthe hydrophile/lipophile balance (HLB) and is a valuable tool incategorizing and selecting surfactants in the preparation offormulations. Surfactants may be classified into different classes basedon the nature of the hydrophilic group: nonionic, anionic, cationic andamphoteric (Rieger, in Pharmaceutical Dosage Forms, Lieberman, Riegerand Banker (Eds.), 1988, Marcel Dekker, Inc., New York, N.Y., volume 1,p. 285).

Naturally occurring emulsifiers used in emulsion formulations includelanolin, beeswax, phosphatides, lecithin and acacia. Absorption basespossess hydrophilic properties such that they can soak up water to formw/o emulsions yet retain their semisolid consistencies, such asanhydrous lanolin and hydrophilic petrolatum. Finely divided solids havealso been used as good emulsifiers especially in combination withsurfactants and in viscous preparations. These include polar inorganicsolids, such as heavy metal hydroxides, nonswelling clays such asbentonite, attapulgite, hectorite, kaolin, montmorillonite, colloidalaluminum silicate and colloidal magnesium aluminum silicate, pigmentsand nonpolar solids such as carbon or glyceryl tristearate.

A large variety of non-emulsifying materials can also be included inemulsion formulations and contribute to the properties of emulsions.These include, but are not limited to, fats, oils, waxes, fatty acids,fatty alcohols, fatty esters, humectants, hydrophilic colloids,preservatives and antioxidants (Block, in Pharmaceutical Dosage Forms,Lieberman, Rieger and Banker (Eds.), 1988, Marcel Dekker, Inc., NewYork, N.Y., volume 1, p. 335; Idson, in Pharmaceutical Dosage Forms,Lieberman, Rieger and Banker (Eds.), 1988, Marcel Dekker, Inc., NewYork, N.Y., volume 1, p. 199).

Hydrophilic colloids or hydrocolloids include naturally occurring gumsand synthetic polymers such as polysaccharides (for example, acacia,agar, alginic acid, carrageenan, guar gum, karaya gum, and tragacanth),cellulose derivatives (for example, carboxymethylcellulose andcarboxypropylcellulose), and synthetic polymers (for example, carbomers,cellulose ethers, and carboxyvinyl polymers). These disperse or swell inwater to form colloidal solutions that stabilize emulsions by formingstrong interfacial films around the dispersed-phase droplets and byincreasing the viscosity of the external phase.

Since emulsions often contain a number of ingredients such ascarbohydrates, proteins, sterols and phosphatides that may readilysupport the growth of microbes, these formulations often incorporatepreservatives. Commonly used preservatives included in emulsionformulations include methyl paraben, propyl paraben, quaternary ammoniumsalts, benzalkonium chloride, esters of p-hydroxybenzoic acid, and boricacid. Antioxidants are also commonly added to emulsion formulations toprevent deterioration of the formulation. Antioxidants used can be freeradical scavengers such as tocopherols, alkyl gallates, butylatedhydroxyanisole, butylated hydroxytoluene, or reducing agents such asascorbic acid and sodium metabisulfite, and antioxidant synergists suchas citric acid, tartaric acid, and lecithin.

The applications of emulsion formulations via dermatological, oral andparenteral routes and methods for their manufacture have been reviewedin the literature (Idson, in Pharmaceutical Dosage Forms, Lieberman,Rieger and Banker (Eds.), 1988, Marcel Dekker, Inc., New York, N.Y.,volume 1, p. 199). Emulsion formulations for oral delivery have beenvery widely used because of ease of formulation, as well as efficacyfrom an absorption and bioavailability standpoint (Rosoff, inPharmaceutical Dosage Forms, Lieberman, Rieger and Banker (Eds.), 1988,Marcel Dekker, Inc., New York, N.Y., volume 1, p. 245; Idson, inPharmaceutical Dosage Forms, Lieberman, Rieger and Banker (Eds.), 1988,Marcel Dekker, Inc., New York, N.Y., volume 1, p. 199).

Exemplary surfactants for inclusion in the particles disclosed hereininclude but are not limited to, ionic surfactants, non-ionicsurfactants, Brij 96, polyoxyethylene oleyl ethers, polyglycerol fattyacid esters, tetraglycerol monolaurate (ML310), tetraglycerol monooleate(MO310), hexaglycerol monooleate (PO310), hexaglycerol pentaoleate(PO500), decaglycerol monocaprate (MCA750), decaglycerol monooleate(MO750), decaglycerol sequioleate (SO750), decaglycerol decaoleate(DAO750), alone or in combination with cosurfactants. The cosurfactant,usually a short-chain alcohol such as ethanol, 1-propanol, and1-butanol, serves to increase the interfacial fluidity by penetratinginto the surfactant film and consequently creating a disordered filmbecause of the void space generated among surfactant molecules.Microemulsions can, however, be prepared without the use ofcosurfactants and alcohol-free self-emulsifying microemulsion systemsare known in the art. The aqueous phase can typically be, but is notlimited to, water, an aqueous solution of the drug, glycerol, PEG300,PEG400, polyglycerols, propylene glycols, and derivatives of ethyleneglycol. The oil phase can include, but is not limited to, materials suchas Captex 300, Captex 355, Capmul MCM, fatty acid esters, medium chain(C8-C12) mono, di, and tri-glycerides, polyoxyethylated glyceryl fattyacid esters, fatty alcohols, polyglycolized glycerides, saturatedpolyglycolized C8-C10 glycerides, vegetable oils and silicone oil.

Microemulsions are particularly of interest from the standpoint of drugsolubilization and the enhanced absorption of drugs. Lipid basedmicroemulsions (both o/w and w/o) have been proposed to enhance the oralbioavailability of drugs, including peptides (see e.g., U.S. Pat. Nos.6,191,105; 7,063,860; 7,070,802; 7,157,099; Constantinides et al.,Pharmaceutical Research, 1994, 11, 1385-1390; Ritschel, Meth. Find. Exp.Clin. Pharmacol., 1993, 13, 205). Microemulsions afford advantages ofimproved drug solubilization, protection of drug from enzymatichydrolysis, possible enhancement of drug absorption due tosurfactant-induced alterations in membrane fluidity and permeability,ease of preparation, ease of oral administration over solid dosageforms, improved clinical potency, and decreased toxicity (see e.g., U.S.Pat. Nos. 6,191,105; 7,063,860; 7,070,802; 7,157,099; Constantinides etal., Pharmaceutical Research, 1994, 11, 1385; Ho et al., J. Pharm. Sci.,1996, 85, 138-143). Often microemulsions can form spontaneously whentheir components are brought together at ambient temperature. This canbe particularly advantageous when formulating thermolabile drugs.Microemulsions have also been effective in the transdermal delivery ofactive components in both cosmetic and pharmaceutical applications. Itis expected that the microemulsion compositions and formulations of thepresent invention will facilitate the increased systemic absorption ofthe platinum based compounds from the gastrointestinal tract, as well asimprove the local cellular uptake of platinum based compounds disclosedherein.

Without wishing to be bound by a theory, nanoparticles disclosed hereinhave higher uptake of platinum in cancer cells relative to cisplatin andoxaliplatin. In some embodiments, the nanoparticles disclosed hereinhave about 25%, about 50%, about 75%, about 1-fold, about 5-folds, about10-folds, about 15-folds, about 20-folds, about 25-folds or higherplatinum uptake in cancer cells relative to cisplatin or oxaliplatin atequivalent dosage.

In addition, the nanoparticles disclosed herein also have higheraccumulation of platinum in tissue, such as, but not limited to a tumor,relative to cisplatin and oxaliplatin when dosed at equivalent amount.For example, the nanoparticles disclosed herein have about 25%, about50%, about 75%, about 1-fold, about 5-folds, about 10-folds, about15-folds, about 20-folds, about 25-folds or higher platinum accumulationtissue relative to cisplatin or oxaliplatin when dosed at equivalentamounts.

The design and synthesis of oxaliplatin nanoparticle is based on theirstructure-activity relationship. The present disclosure describes thesynthesis of various platinum based amphiphiles by functional groupinterchange chemistry. In a specific embodiment, for obtaining acarbamate linkage, cholesteryl chloroformate is employed as a startingmaterial to which ethylene diamine (a) (linker) is added to obtain anethylene diamine conjugated cholesterol where one amine group ofethylene diamine forms a carbamate bond with cholesterol and the otheramine is free (FIG. 1A). In the next step, the free amine reacts withone of the carboxyl groups of succinic anhydride (b) (a dicarbonylderivative which is capable of forming seven membered ring molecules) toform an amide bond and the other free carboxylic acid group remains forplatinum co-ordination (FIG. 1A). Dichlorodiamino-platinum (II) [RRisomer] is hydrated with silver nitrate overnight to obtain aquatedoxaliplatin (FIG. 1B) which forms an adduct with intermediate II/III/IV(as depicted in FIG. 1A) via. the formation of a covalentmonocarboxylato bond and another co-ordination bond of amide oxygen(FIG. 1C).

Similarly, in yet another embodiment, the synthesis of ether linkedplatinum amphiphiles have been summarized in FIG. 2 wherein, cholesterolmolecule is initially transformed into intermediate I by functionalgroup interchange (FIG. 2A). This intermediate I is transformed intointermediate II which in turn leads to final adduct by similar reactionsteps as used for the synthesis of carbamate linkage as described above.Similarly, for synthesizing the six and five membered ring molecules asdescribed above, monoethyl esters of malonic acid and oxalic acid havebeen used for the synthesis of both carbamate linked and ether linkedplatinum amphiphiles respectively (FIGS. 1A and 2A) followed by ethylester deprotection.

After the synthesis, the final platinum adducts are formulated intonanoparticles with different co-lipids selected from Soy-PC, DOPE, DOPCetc and stabilizers selected from DSPE-PEG-OMe etc. Further, thecharacterization of all intermediates are performed by ¹HNMR and thecharacterization of the final oxaliplatin amphiphile molecule is carriedout using ¹HNMR and MALDI-TOF respectively.

TABLE 1 Classification of Platinum (II) compounds (includes Formula IIIcompounds) based upon coordination environment: Class-III Symmetriccoordination with one side DACH and other side O—O coordination, but theasymmetry is introduced by secondary atom/s Class-I Class-II which isnot Class-IV Symmetric Asymmetric connected to Pt Symmetric coordinationwith coordination with [a sub class of coordination with one side DACHone side DACH Class - I (a)] no DACH

Class-V Asymmetric coordination with no DACH a) ′Pt′ is connected a)′PT′ is a) One ′O′ a) One side two to two ′O′ connected to ′P′ connectedto ′S′ ′O ′other side Compounds 52, 55, and ′O′ other ′O′ connected two′Cl′ 65, 74, 75, 76, 79 Compounds 46, to ′CO′ Compound 58 49 Compounds63, 69, 70 b) ′Pt′ is connected b) Pt′ is b) One ′O′ b) One side two b)With to two ′P′ connected to ′P′ connected to ′C═O′ ′S′ other sideterpyridine Compounds 43, 44, and ′S′ other ′O′ connected two ′Cl′ groupwith 45 Compounds 47, to ′C═C′ Compound 59 one ′O′ 50 Compounds 66, 67,coordination 68 Compound 95 c) ′Pt′ is connected c) ′Pt′ is c) One ′O′c) One side two to two ′S′ connected to ′P′ connected to ′C═O′ ′S′ otherside Compounds 53, 56, and ′Se′ other ′′O′ connected two ′O′ 64Compounds 48, to ′P′ Compound 61 51 Compound 71 d) ′Pt′ is connected d)′Pt′ is d) One ′O′ d) One side two to two ′Se′ connected to ′O′connected to ′C═O′ ′Se′ other side Compounds 54, 57 and ′S′ other ′O′connected two ′Cl′ Compounds 73, to ′N′ Compound 60 82, 84, 85 Compound72 e) ′Pt′ is e) One ′O′ e) One side two connected to ′O′ connected to′S′ ′Se′ other side and ′Cl′ other ′O′ connected two ′O′ Compounds 77,to ′C′ Compound 62 78, 81 Compound 80 f) ′Pt′ is connected to ′S′ and′Cl′ Compound 83

Without wishing to be bound by a theory, the nanoparticle compositionsof the present disclosure show significant cancer cell killing efficacy.Exemplary nanoparticles were tested in different cancer cell lines andit was observed that the compounds demonstrated significantly bettercell killing efficacy than the control compounds such as conventionallyknown platinum drugs oxaliplatin, cisplatin, oxaliplatin, carboplatin,paraplatin and sartraplatin.

Accordingly, in another aspect, described herein is a method of treatingcancer, Generally, the method comprises administering a therapeuticallyeffective amount of a platinum based compounds disclosed herein to asubject in need thereof.

The phrase “therapeutically-effective amount” as used herein means thatamount of a compound, material, or composition comprising a compound ofthe present invention which is effective for producing some desiredtherapeutic effect in at least a sub-population of cells in an animal ata reasonable benefit/risk ratio applicable to any medical treatment.Determination of a therapeutically effective amount is well within thecapability of those skilled in the art. Generally, a therapeuticallyeffective amount can vary with the subject's history, age, condition,sex, as well as the severity and type of the medical condition in thesubject, and administration of other agents alleviate the disease ordisorder to be treated.

Usually the amount of active compounds is between 0.1-95% by weight ofthe preparation, preferably between 0.2-20% by weight in preparationsfor parenteral use and preferably between 1 and 50% by weight inpreparations for oral administration.

Toxicity and therapeutic efficacy can be determined by standardpharmaceutical procedures in cell cultures or experimental animals,e.g., for determining the LD50 (the dose lethal to 50% of thepopulation) and the ED50 (the dose therapeutically effective in 50% ofthe population). The dose ratio between toxic and therapeutic effects isthe therapeutic index and it can be expressed as the ratio LD50/ED50.Compositions that exhibit large therapeutic indices are preferred. Asused herein, the term ED denotes effective dose and is used inconnection with animal models. The term EC denotes effectiveconcentration and is used in connection with in vitro models.

The data obtained from the cell culture assays and animal studies can beused in formulating a range of dosage for use in humans. The dosage ofsuch compounds lies preferably within a range of circulatingconcentrations that include the ED50 with little or no toxicity. Thedosage can vary within this range depending upon the dosage formemployed and the route of administration utilized.

The therapeutically effective dose can be estimated initially from cellculture assays. A dose can be formulated in animal models to achieve acirculating plasma concentration range that includes the IC50 (i.e., theconcentration of the therapeutic which achieves a half-maximalinhibition of symptoms) as determined in cell culture. Levels in plasmacan be measured, for example, by high performance liquid chromatography.The effects of any particular dosage can be monitored by a suitablebioassay.

The dosage can be determined by a physician and adjusted, as necessary,to suit observed effects of the treatment. Generally, the compositionsare administered so that the agent is given at a dose from 1 μg/kg to150 mg/kg, 1 μg/kg to 100 mg/kg, 1 μg/kg to 50 mg/kg, 1 μg/kg to 20mg/kg, 1 μg/kg to 10 mg/kg, 1 μg/kg to 1 mg/kg, 100 μg/kg to 100 mg/kg,100 μg/kg to 50 mg/kg, 100 μg/kg to 20 mg/kg, 100 μg/kg to 10 mg/kg, 100μg/kg to 1 mg/kg, 1 mg/kg to 100 mg/kg, 1 mg/kg to 50 mg/kg, 1 mg/kg to20 mg/kg, 1 mg/kg to 10 mg/kg, 10 mg/kg to 100 mg/kg, 10 mg/kg to 50mg/kg, or 10 mg/kg to 20 mg/kg. It is to be understood that ranges givenhere include all intermediate ranges, for example, the range 1 mg/kg to10 mg/kg includes 1 mg/kg to 2 mg/kg, 1 mg/kg to 3 mg/kg, 1 mg/kg to 4mg/kg, 1 mg/kg to 5 mg/kg, 1 mg/kg to 6 mg/kg, 1 mg/kg to 7 mg/kg, 1mg/kg to 8 mg/kg, 1 mg/kg to 9 mg/kg, 2 mg/kg to 10 mg/kg, 3 mg/kg to 10mg/kg, 4 mg/kg to 10 mg/kg, 5 mg/kg to 10 mg/kg, 6 mg/kg to 10 mg/kg, 7mg/kg to 10 mg/kg, 8 mg/kg to 10 mg/kg, 9 mg/kg to 10 mg/kg, and thelike. It is to be further understood that the ranges intermediate to thegiven above are also within the scope of this invention, for example, inthe range 1 mg/kg to 10 mg/kg, dose ranges such as 2 mg/kg to 8 mg/kg, 3mg/kg to 7 mg/kg, 4 mg/kg to 6 mg/kg, and the like.

In some embodiments, the compositions are administered at a dosage sothat the agent has an in vivo concentration of less than 500 nM, lessthan 400 nM, less than 300 nM, less than 250 nM, less than 200 nM, lessthan 150 nM, less than 100 nM, less than 50 nM, less than 25 nM, lessthan 20, nM, less than 10 nM, less than 5 nM, less than 1 nM, less than0.5 nM, less than 0.1 nM, less than 0.05, less than 0.01, nM, less than0.005 nM, less than 0.001 nM after 15 mins, 30 mins, 1 hr, 1.5 hrs, 2hrs, 2.5 hrs, 3 hrs, 4 hrs, 5 hrs, 6 hrs, 7 hrs, 8 hrs, 9 hrs, 10 hrs,11 hrs, 12 hrs or more of time of administration.

With respect to duration and frequency of treatment, it is typical forskilled clinicians to monitor subjects in order to determine when thetreatment is providing therapeutic benefit, and to determine whether toincrease or decrease dosage, increase or decrease administrationfrequency, discontinue treatment, resume treatment or make otheralteration to treatment regimen. The dosing schedule can vary from oncea week to daily depending on a number of clinical factors, such as thesubject's sensitivity to the polypeptides. The desired dose can beadministered every day or every third, fourth, fifth, or sixth day. Thedesired dose can be administered at one time or divided into subdoses,e.g., 2-4 subdoses and administered over a period of time, e.g., atappropriate intervals through the day or other appropriate schedule.Such sub-doses can be administered as unit dosage forms. In someembodiments of the aspects described herein, administration is chronic,e.g., one or more doses daily over a period of weeks or months. Examplesof dosing schedules are administration daily, twice daily, three timesdaily or four or more times daily over a period of 1 week, 2 weeks, 3weeks, 4 weeks, 1 month, 2 months, 3 months, 4 months, 5 months, or 6months or more.

In some embodiments, the platinum based compound can be administrated toa subject in combination with a pharmaceutically active agent, e.g., asecond therapeutic agent. Exemplary pharmaceutically active compoundinclude, but are not limited to, those found in Harrison's Principles ofInternal Medicine, 13^(th) Edition, Eds. T. R. Harrison et al.McGraw-Hill N.Y., NY; Physicians Desk Reference, 50^(th) Edition, 1997,Oradell N.J., Medical Economics Co.; Pharmacological Basis ofTherapeutics, 8^(th) Edition, Goodman and Gilman, 1990; and UnitedStates Pharmacopeia, The National Formulary, USP XII NF XVII, 1990, thecomplete contents of all of which are incorporated herein by reference.The platinum based compound and the second therapeutic agent can beadministrated to the subject in the same pharmaceutical composition orin different pharmaceutical compositions (at the same time or atdifferent times).

As used herein, the term “administer” refers to the placement of acomposition into a subject by a method or route which results in atleast partial localization of the composition at a desired site suchthat desired effect is produced. A compound or composition describedherein can be administered by any appropriate route known in the artincluding, but not limited to, oral or parenteral routes, includingintravenous, intramuscular, subcutaneous, transdermal, airway (aerosol),pulmonary, nasal, rectal, and topical (including buccal and sublingual)administration.

Exemplary modes of administration include, but are not limited to,injection, infusion, instillation, inhalation, or ingestion. “Injection”includes, without limitation, intravenous, intramuscular, intraarterial,intrathecal, intraventricular, intracapsular, intraorbital,intracardiac, intradermal, intraperitoneal, transtracheal, subcutaneous,subcuticular, intraarticular, sub capsular, subarachnoid, intraspinal,intracerebro spinal, and intrasternal injection and infusion. In someembodiments, the compositions are administered by intravenous infusionor injection.

As used herein, the term “cancer” refers to an uncontrolled growth ofcells that may interfere with the normal functioning of the bodilyorgans and systems. Cancers that migrate from their original locationand seed vital organs can eventually lead to the death of the subjectthrough the functional deterioration of the affected organs. Metastasisis a cancer cell or group of cancer cells, distinct from the primarytumor location resulting from the dissemination of cancer cells from theprimary tumor to other parts of the body. At the time of diagnosis ofthe primary tumor mass, the subject may be monitored for the presence ofin transit metastases, e.g., cancer cells in the process ofdissemination. As used herein, the term cancer, includes, but is notlimited to the following types of cancer, breast cancer, biliary tractcancer, bladder cancer, brain cancer including glioblastomas andmedulloblastomas; cervical cancer; choriocarcinoma; colon cancer;endometrial cancer; esophageal cancer, gastric cancer; hematologicalneoplasms including acute lymphocytic and myelogenous leukemia; T-cellacute lymphoblastic leukemia/lymphoma; hairy cell leukemia; chronicmyelogenous leukemia, multiple myeloma; AIDS-associated leukemias andadult T-cell leukemia lymphoma; intraepithelial neoplasms includingBowen's disease and Paget's disease; liver cancer; lung cancer;lymphomas including Hodgkin's disease and lymphocytic lymphomas;neuroblastomas; oral cancer including squamous cell carcinoma; ovariancancer including those arising from epithelial cells, stromal cells,germ cells and mesenchymal cells; pancreatic cancer; prostate cancer;rectal cancer; sarcomas including leiomyosarcoma, rhabdomyosarcoma,liposarcoma, fibrosarcoma, and osteosarcoma; skin cancer includingmelanoma, Merkel cell carcinoma, Kaposi's sarcoma, basal cell carcinoma,and squamous cell cancer; testicular cancer including germinal tumorssuch as seminoma, non-seminoma (teratomas, choriocarcinomas), stromaltumors, and germ cell tumors; thyroid cancer including thyroidadenocarcinoma and medullar carcinoma; and renal cancer includingadenocarcinoma, Wilms tumor. Examples of cancer include but are notlimited to, carcinoma, including adenocarcinoma, lymphoma, blastoma,melanoma, sarcoma, and leukemia. More particular examples of suchcancers include squamous cell cancer, small-cell lung cancer, non-smallcell lung cancer, gastrointestinal cancer, Hodgkin's and non-Hodgkin'slymphoma, pancreatic cancer, Glioblastoma, cervical cancer, ovariancancer, liver cancer such as hepatic carcinoma and hepatoma, bladdercancer, breast cancer, colon cancer, colorectal cancer, endometrialcarcinoma, salivary gland carcinoma, kidney cancer such as renal cellcarcinoma and Wilms' tumors, basal cell carcinoma, melanoma, prostatecancer, vulval cancer, thyroid cancer, testicular cancer, esophagealcancer, and various types of head and neck cancer. Other cancers will beknown to the artisan.

As used herein, the term “cancer” includes, but is not limited to, solidtumors and blood born tumors. The term cancer refers to disease of skin,tissues, organs, bone, cartilage, blood and vessels. The term “cancer”further encompasses primary and metastatic cancers. Examples of cancersthat can be treated with the compounds of the invention include, but arenot limited to, carcinoma, including that of the bladder, breast, colon,kidney, lung, ovary, pancreas, stomach, cervix, thyroid, and skin,including squamous cell carcinoma; hematopoietic tumors of lymphoidlineage, including, but not limited to, leukemia, acute lymphocyticleukemia, acute lymphoblastic leukemia, B-cell lymphoma, T-celllymphoma, Hodgkin's lymphoma, non-Hodgkin's lymphoma, hairy celllymphoma, and Burkett's lymphoma; hematopoietic tumors of myeloidlineage including, but not limited to, acute and chronic myelogenousleukemias and promyelocytic leukemia; tumors of mesenchymal originincluding, but not limited to, fibrosarcoma, rhabdomyosarcoma, andosteosarcoma; other tumors including melanoma, seminoma,tetratocarcinoma, neuroblastoma, and glioma; tumors of the central andperipheral nervous system including, but not limited to, astrocytoma,neuroblastoma, glioma, and schwannomas; and other tumors including, butnot limited to, xenoderma, pigmentosum, keratoactanthoma, thyroidfollicular cancer, and teratocarcinoma. The methods disclosed herein areuseful for treating patients who have been previously treated forcancer, as well as those who have not previously been treated forcancer. Indeed, the methods and compositions of this invention can beused in first-line and second-line cancer treatments.

As used herein, the term “precancerous condition” has its ordinarymeaning, i.e., an unregulated growth without metastasis, and includesvarious forms of hyperplasia and benign hypertrophy. Accordingly, a“precancerous condition” is a disease, syndrome, or finding that, ifleft untreated, can lead to cancer. It is a generalized state associatedwith a significantly increased risk of cancer. Premalignant lesion is amorphologically altered tissue in which cancer is more likely to occurthan its apparently normal counterpart. Examples of pre-malignantconditions include, but are not limited to, oral leukoplakia, actinickeratosis (solar keratosis), Barrett's esophagus, atrophic gastritis,benign hyperplasia of the prostate, precancerous polyps of the colon orrectum, gastric epithelial dysplasia, adenomatous dysplasia, hereditarynonpolyposis colon cancer syndrome (HNPCC), Barrett's esophagus, bladderdysplasia, precancerous cervical conditions, and cervical dysplasia.

In some embodiments, the cancer is selected from the group consistingof: breast cancer; ovarian cancer; glioma; gastrointestinal cancer;prostate cancer; carcinoma, lung carcinoma, hepatocellular carcinoma,testicular cancer; cervical cancer; endometrial cancer; bladder cancer;head and neck cancer; lung cancer; gastro-esophageal cancer, andgynecological cancer.

In some embodiments, the methods described herein relate to treating asubject having or diagnosed as having cancer. Subjects having cancer canbe identified by a physician using current methods of diagnosing cancer.Symptoms and/or complications of cancer which characterize theseconditions and aid in diagnosis are well known in the art and includebut are not limited to, growth of a tumor, impaired function of theorgan or tissue harboring cancer cells, etc. Tests that may aid in adiagnosis of, e.g. cancer include, but are not limited to, tissuebiopsies and histological examination. A family history of cancer, orexposure to risk factors for cancer (e.g. tobacco products, radiation,etc.) can also aid in determining if a subject is likely to have canceror in making a diagnosis of cancer.

In some embodiments, the method further comprises co-administering oneor more additional anti-cancer therapy to the patient. In someembodiments, the additional therapy is selected from the groupconsisting of surgery, chemotherapy, radiation therapy, thermotherapy,immunotherapy, hormone therapy, laser therapy, anti-angiogenic therapy,and any combinations thereof. In some embodiments, the additionaltherapy comprises administering an anti-cancer agent to the patient. Insome embodiments, the method comprises co-administering the conjugateand an anti-cancer agent or chemotherapeutic agent to the subject. Asused herein, the term “anti-cancer agent” is refers to any compound(including its analogs, derivatives, prodrugs and pharmaceuticallysalts) or composition which can be used to treat cancer. Anti-cancercompounds for use in the present invention include, but are not limitedto, inhibitors of topoisomerase I and II, alkylating agents, microtubuleinhibitors (e.g., taxol), and angiogenesis inhibitors. Exemplaryanti-cancer compounds include, but are not limited to, paclitaxel(taxol); docetaxel; germicitibine; Aldesleukin; Alemtuzumab;alitretinoin; allopurinol; altretamine; amifostine; anastrozole; arsenictrioxide; asparaginase; BCG Live; bexarotene capsules; bexarotene gel;bleomycin; busulfan intravenous; busulfanoral; calusterone;capecitabine; platinate; carmustine; carmustine with Polifeprosanimplant; celecoxib; chlorambucil; cladribine; cyclophosphamide;cytarabine; cytarabine liposomal; dacarbazine; dactinomycin; actinomycinD; Darbepoetin alfa; daunorubicin liposomal; daunorubicin, daunomycin;Denileukin diftitox, dexrazoxane; docetaxel; doxorubicin; doxorubicinliposomal; Dromostanolone propionate; Elliott's B Solution; epirubicin;Epoetin alfa estramustine; etoposide phosphate; etoposide (VP-16);exemestane; Filgrastim; floxuridine (intraarterial); fludarabine;fluorouracil (5-FU); fulvestrant; gemtuzumab ozogamicin; goserelinacetate; hydroxyurea; Ibritumomab Tiuxetan; idarubicin; ifosfamide;imatinib mesylate; interferon alfa-2a; Interferon alfa-2b; irinotecan;letrozole; leucovorin; levamisole; lomustine (CCNU); mechlorethamine(nitrogen mustard); megestrol acetate; melphalan (L-PAM); mercaptopurine(6-MP); mesna; methotrexate; methoxsalen; mitomycin C; mitotane;mitoxantrone; nandrolone phenpropionate; Nofetumomab; LOddC; Oprelvekin;pamidronate; pegademase; Pegaspargase; Pegfilgrastim; pentostatin;pipobroman; plicamycin; mithramycin; porfimer sodium; procarbazine;quinacrine; Rasburicase; Rituximab; Sargramostim; streptozocin;talbuvidine (LDT); talc; tamoxifen; temozolomide; teniposide (VM-26);testolactone; thioguanine (6-TG); thiotepa; topotecan; toremifene;Tositumomab; Trastuzumab; tretinoin (ATRA); Uracil Mustard; valrubicin;valtorcitabine (monoval LDC); vinblastine; vinorelbine; zoledronate; andany mixtures thereof. In some embodiments, the anti-cancer agent is apaclitaxel-carbohydrate conjugate, e.g., a paclitaxel-glucose conjugate,as described in U.S. Pat. No. 6,218,367, content of which is hereinincorporated by reference in its entirety.

The methods of the invention are especially useful in combination withanti-cancer treatments that involve administering a second drug thatacts in a different phase of the cell cycle.

For administration to a subject, the platinum based compounds and/orparticles comprising said platinum based compounds are provided inpharmaceutically acceptable compositions. Accordingly, the disclosurealso provides pharmaceutical compositions comprising the platinum basedcompounds or particles as disclosed herein. These pharmaceuticallyacceptable compositions comprise a therapeutically-effective amount ofone or more of the platinum based compounds or particles describedherein, formulated together with one or more pharmaceutically acceptablecarriers (additives) and/or diluents. The said pharmaceuticalcompositions of the present invention are specially formulated foradministration in solid or liquid form, including those adapted for thefollowing: (1) oral administration, for example, drenches (aqueous ornon-aqueous solutions or suspensions), lozenges, dragees, capsules,pills, tablets (e.g., those targeted for buccal, sublingual, andsystemic absorption), boluses, powders, granules, pastes for applicationto the tongue; (2) parenteral administration, for example, bysubcutaneous, intramuscular, intravenous or epidural injection as, forexample, a sterile solution or suspension, or sustained-releaseformulation; (3) topical application, for example, as a cream, ointment,or a controlled-release patch or spray applied to the skin; (4)intravaginally or intrarectally, for example, as a pessary, cream orfoam; (5) sublingually; (6) ocularly; (7) transdermally; (8)transmucosally; or (9) nasally. Additionally, the compounds of thepresent disclosure can be implanted into a patient or injected using adrug delivery system.

As used herein, the term “pharmaceutically acceptable” refers to thosecompounds, materials, compositions, and/or dosage forms which are,within the scope of sound medical judgment, suitable for use in contactwith the tissues of human beings and animals without excessive toxicity,irritation, allergic response, or other problem or complication,commensurate with a reasonable benefit/risk ratio.

As used herein, the term “pharmaceutically-acceptable carrier” means apharmaceutically-acceptable material, composition or vehicle, such as aliquid or solid filler, diluent, excipient, manufacturing aid (e.g.,lubricant, talc magnesium, calcium or zinc stearate, or steric acid), orsolvent encapsulating material, involved in carrying or transporting thesubject compound from one organ, or portion of the body, to anotherorgan, or portion of the body. Each carrier must be “acceptable” in thesense of being compatible with the other ingredients of the formulationand not injurious to the patient. Some examples of materials which canserve as pharmaceutically-acceptable carriers include: (1) sugars, suchas lactose, glucose and sucrose; (2) starches, such as corn starch andpotato starch; (3) cellulose, and its derivatives, such as sodiumcarboxymethyl cellulose, methylcellulose, ethyl cellulose,microcrystalline cellulose and cellulose acetate; (4) powderedtragacanth; (5) malt; (6) gelatin; (7) lubricating agents, such asmagnesium stearate, sodium lauryl sulfate and talc; (S) excipients, suchas cocoa butter and suppository waxes; (9) oils, such as peanut oil,cottonseed oil, safflower oil, sesame oil, olive oil, corn oil andsoybean oil; (10) glycols, such as propylene glycol; (11) polyols, suchas glycerin, sorbitol, mannitol and polyethylene glycol (PEG); (12)esters, such as ethyl oleate and ethyllaurate; (13) agar; (14) bufferingagents, such as magnesium hydroxide and aluminum hydroxide; (15) alginicacid; (16) pyrogen-free water; (17) isotonic saline; (IS) Ringer'ssolution; (19) ethyl alcohol; (20) pH buffered solutions; (21)polyesters, polycarbonates and/or polyanhydrides; (22) bulking agents,such as polypeptides and amino acids (23) serum component, such as serumalbumin, HDL and LDL; (22) C2-C12 alcohols, such as ethanol; and (23)other non-toxic compatible substances employed in pharmaceuticalformulations. Wetting agents, coloring agents, release agents, coatingagents, sweetening agents, flavoring agents, perfuming agents,preservative and antioxidants can also be present in the formulation.The terms such as “excipient”, “carrier”, “pharmaceutically acceptablecarrier” or the likes are used interchangeably herein.

In some embodiments, the pharmaceutical composition comprising aplatinum based compound can be a parenteral dose form. Sinceadministration of parenteral dosage forms typically bypasses thepatient's natural defenses against contaminants, parenteral dosage formsare preferably sterile or capable of being sterilized prior toadministration to a patient. Examples of parenteral dosage formsinclude, but are not limited to, solutions ready for injection, dryproducts ready to be dissolved or suspended in a pharmaceuticallyacceptable vehicle for injection, suspensions ready for injection, andemulsions. In addition, controlled-release parenteral dosage forms canbe prepared for administration of a patient, including, but not limitedto, DUROS®-type dosage forms and dose-dumping.

Suitable vehicles that can be used to provide parenteral dosage forms ofa composition as described herein are well known to those skilled in theart. Examples include, without limitation: sterile water; water forinjection USP; saline solution; glucose solution; aqueous vehicles suchas but not limited to, sodium chloride injection, Ringer's injection,dextrose Injection, dextrose and sodium chloride injection, and lactatedRinger's injection; water-miscible vehicles such as, but not limited to,ethyl alcohol, polyethylene glycol, and propylene glycol; andnon-aqueous vehicles such as, but not limited to, corn oil, cottonseedoil, peanut oil, sesame oil, ethyl oleate, isopropyl myristate, andbenzyl benzoate. Compounds that alter or modify the solubility of apharmaceutically acceptable salt can also be incorporated into theparenteral dosage forms of the disclosure, including conventional andcontrolled-release parenteral dosage forms.

Pharmaceutical compositions can also be formulated to be suitable fororal administration, for example as discrete dosage forms, such as, butnot limited to, tablets (including without limitation scored or coatedtablets), pills, caplets, capsules, chewable tablets, powder packets,cachets, troches, wafers, aerosol sprays, or liquids, such as but notlimited to, syrups, elixirs, solutions or suspensions in an aqueousliquid, a non-aqueous liquid, an oil-in-water emulsion, or awater-in-oil emulsion. Such compositions contain a predetermined amountof the pharmaceutically acceptable salt of the disclosed compounds, andmay be prepared by methods of pharmacy well known to those skilled inthe art. See generally, Remington: The Science and Practice of Pharmacy,21st Ed., Lippincott, Williams, and Wilkins, Philadelphia Pa. (2005).

Conventional dosage forms generally provide rapid or immediate drugrelease from the formulation. Depending on the pharmacology andpharmacokinetics of the drug, use of conventional dosage forms can leadto wide fluctuations in the concentrations of the drug in a patient'sblood and other tissues. These fluctuations can impact a number ofparameters, such as dose frequency, onset of action, duration ofefficacy, maintenance of therapeutic blood levels, toxicity, sideeffects, and the like. Advantageously, controlled-release formulationscan be used to control a drug's onset of action, duration of action,plasma levels within the therapeutic window, and peak blood levels. Inparticular, controlled- or extended-release dosage forms or formulationscan be used to ensure that the maximum effectiveness of a drug isachieved while minimizing potential adverse effects and safety concerns,which can occur both from under-dosing a drug (i.e., going below theminimum therapeutic levels) as well as exceeding the toxicity level forthe drug. In some embodiments, a composition as described herein can beadministered in a sustained release formulation.

Controlled-release pharmaceutical products have a common goal ofimproving drug therapy over that achieved by their non-controlledrelease counterparts. Ideally, the use of an optimally designedcontrolled-release preparation in medical treatment is characterized bya minimum of drug substance being employed to cure or control thecondition in a minimum amount of time. Advantages of controlled-releaseformulations include: 1) extended activity of the drug; 2) reduceddosage frequency; 3) increased patient compliance; 4) usage of lesstotal drug; 5) reduction in local or systemic side effects; 6)minimization of drug accumulation; 7) reduction in blood levelfluctuations; 8) improvement in efficacy of treatment; 9) reduction ofpotentiation or loss of drug activity; and 10) improvement in speed ofcontrol of diseases or conditions. Kim, Cherng-ju, Controlled ReleaseDosage Form Design, 2 (Technomic Publishing, Lancaster, Pa.: 2000).

Most controlled-release formulations are designed to initially releasean amount of drug (active ingredient) that promptly produces the desiredtherapeutic effect, and gradually and continually release other amountsof drug to maintain this level of therapeutic or prophylactic effectover an extended period of time. In order to maintain this constantlevel of drug in the body, the drug must be released from the dosageform at a rate that will replace the amount of drug being metabolizedand excreted from the body. Controlled-release of an active ingredientcan be stimulated by various conditions including, but not limited to,pH, ionic strength, osmotic pressure, temperature, enzymes, water, andother physiological conditions or compounds.

A variety of known controlled- or extended-release dosage forms,formulations, and devices can be adapted for use with the salts andcompositions of the disclosure. Examples include, but are not limitedto, those described in U.S. Pat. Nos. 3,845,770; 3,916,899; 3,536,809;3,598,123; 4,008,719; 5,674,533; 5,059,595; 5,591,767; 5,120,548;5,073,543; 5,639,476; 5,354,556; 5,733,566; and 6,365,185 B1; each ofwhich is incorporated herein by reference. These dosage forms can beused to provide slow or controlled-release of one or more activeingredients using, for example, hydroxypropylmethyl cellulose, otherpolymer matrices, gels, permeable membranes, osmotic systems (such asOROS® (Alza Corporation, Mountain View, Calif. USA)), or a combinationthereof to provide the desired release profile in varying proportions.

Some Selected Definitions

For convenience, certain terms employed herein, in the specification,examples and appended claims are collected herein. Unless statedotherwise, or implicit from context, the following terms and phrasesinclude the meanings provided below. Unless explicitly stated otherwise,or apparent from context, the terms and phrases below do not exclude themeaning that the term or phrase has acquired in the art to which itpertains. The definitions are provided to aid in describing particularembodiments, and are not intended to limit the claimed invention,because the scope of the invention is limited only by the claims.Further, unless otherwise required by context, singular terms shallinclude pluralities and plural terms shall include the singular.

Unless defined otherwise, all technical and scientific terms used hereinhave the same meaning as those commonly understood to one of ordinaryskill in the art to which this invention pertains. Although any knownmethods, devices, and materials may be used in the practice or testingof the invention, the methods, devices, and materials in this regard aredescribed herein.

As used herein the term “comprising” or “comprises” is used in referenceto compositions, methods, and respective component(s) thereof, that areessential to the invention, yet open to the inclusion of unspecifiedelements, whether essential or not.

The singular terms “a,” “an,” and “the” include plural referents unlesscontext clearly indicates otherwise. Similarly, the word “or” isintended to include “and” unless the context clearly indicatesotherwise.

Other than in the operating examples, or where otherwise indicated, allnumbers expressing quantities of ingredients or reaction conditions usedherein should be understood as modified in all instances by the term“about.” The term “about” when used in connection with percentages maymean 5% of the value being referred to. For example, about 100 meansfrom 95 to 105.

Although methods and materials similar or equivalent to those describedherein can be used in the practice or testing of this disclosure,suitable methods and materials are described below. The term “comprises”means “includes.” The abbreviation, “e.g.” is derived from the Latinexempli gratia, and is used herein to indicate a non-limiting example.Thus, the abbreviation “e.g.” is synonymous with the term “for example.”

The terms “decrease”, “reduced”, “reduction”, “decrease” or “inhibit”are all used herein generally to mean a decrease by a statisticallysignificant amount. However, for avoidance of doubt, “reduced”,“reduction” or “decrease” or “inhibit” means a decrease by at least 10%as compared to a reference level, for example a decrease by at leastabout 20%, or at least about 30%, or at least about 40%, or at leastabout 50%, or at least about 60%, or at least about 70%, or at leastabout 80%, or at least about 90% or up to and including a 100% decrease(e.g. absent level as compared to a reference sample), or any decreasebetween 10-100% as compared to a reference level.

The terms “increased”, “increase” or “enhance” or “activate” are allused herein to generally mean an increase by a statically significantamount; for the avoidance of any doubt, the terms “increased”,“increase” or “enhance” or “activate” means an increase of at least 10%as compared to a reference level, for example an increase of at leastabout 20%, or at least about 30%, or at least about 40%, or at leastabout 50%, or at least about 60%, or at least about 70%, or at leastabout 80%, or at least about 90% or up to and including a 100% increaseor any increase between 10-100% as compared to a reference level, or atleast about a 2-fold, or at least about a 3-fold, or at least about a4-fold, or at least about a 5-fold or at least about a 10-fold increase,or any increase between 2-fold and 10-fold or greater as compared to areference level.

The term “statistically significant” or “significantly” refers tostatistical significance and generally means at least two standarddeviation (2SD) away from a reference level. The term refers tostatistical evidence that there is a difference. It is defined as theprobability of making a decision to reject the null hypothesis when thenull hypothesis is actually true.

As used herein, the terms “treat,” “treatment,” “treating,” or“amelioration” refer to therapeutic treatments, wherein the object is toreverse, alleviate, ameliorate, inhibit, slow down or stop theprogression or severity of a condition associated with a disease ordisorder, e.g. cancer. The term “treating” includes reducing oralleviating at least one adverse effect or symptom of a condition,disease or disorder associated with a cancer. Treatment is generally“effective” if one or more symptoms or clinical markers are reduced.Alternatively, treatment is “effective” if the progression of a diseaseis reduced or halted. That is, “treatment” includes not just theimprovement of symptoms or markers, but also a cessation of, or at leastslowing of, progress or worsening of symptoms compared to what would beexpected in the absence of treatment. Beneficial or desired clinicalresults include, but are not limited to, alleviation of one or moresymptom(s), diminishment of extent of disease, stabilized (i.e., notworsening) state of disease, delay or slowing of disease progression,amelioration or palliation of the disease state, remission (whetherpartial or total), and/or decreased mortality, whether detectable orundetectable. The term “treatment” of a disease also includes providingrelief from the symptoms or side-effects of the disease (includingpalliative treatment).

As used herein, “management” or “managing” refers to preventing adisease or disorder from occurring in a subject, decreasing the risk ofdeath due to a disease or disorder, delaying the onset of a disease ordisorder, inhibiting the progression of a disease or disorder, partialor complete cure of a disease or disorder and/or adverse effectattributable to the said disease or disorder, obtaining a desiredpharmacologic and/or physiologic effect (the effect may be prophylacticin terms of completely or partially preventing a disorder or disease orcondition, or a symptom thereof and/or may be therapeutic in terms of apartial or complete cure for a disease or disorder and/or adverse effectattributable to the disease or disorder), relieving a disease ordisorder (i.e. causing regression of the disease or disorder). Further,the present disclosure also envisages treating the said disease byadministering the therapeutic composition of the instant disclosure.

The terms “subject” and “individual” are used interchangeably herein,and mean a human or animal. Usually the animal is a vertebrate such as aprimate, rodent, domestic animal or game animal. Primates includechimpanzees, cynomologous monkeys, spider monkeys, and macaques, e.g.,Rhesus. Rodents include mice, rats, woodchucks, ferrets, rabbits andhamsters. Domestic and game animals include cows, horses, pigs, deer,bison, buffalo, feline species, e.g., domestic cat, canine species,e.g., dog, fox, wolf, avian species, e.g., chicken, emu, ostrich, andfish, e.g., trout, catfish and salmon. Patient or subject includes anysubset of the foregoing, e.g., all of the above, but excluding one ormore groups or species such as humans, primates or rodents. In certainembodiments, the subject is a mammal, e.g., a primate, e.g., a human.The terms, “patient” and “subject” are used interchangeably herein. Theterms, “patient” and “subject” are used interchangeably herein.

Preferably, the subject is a mammal. The mammal can be a human,non-human primate, mouse, rat, dog, cat, horse, or cow, but are notlimited to these examples. Mammals other than humans can beadvantageously used as subjects that represent animal models of cancer.In addition, the methods described herein can be used to treatdomesticated animals and/or pets. A subject can be male or female. Asubject can be one who has been previously diagnosed with or identifiedas suffering from cancer, but need not have already undergone treatment.

The description of embodiments of the disclosure is not intended to beexhaustive or to limit the disclosure to the precise form disclosed.While specific embodiments of, and examples for, the disclosure aredescribed herein for illustrative purposes, various equivalentmodifications are possible within the scope of the disclosure, as thoseskilled in the relevant art will recognize. For example, while methodsteps or functions are presented in a given order, alternativeembodiments may perform functions in a different order, or functions maybe performed substantially concurrently. The teachings of the disclosureprovided herein can be applied to other procedures or methods asappropriate. The various embodiments described herein can be combined toprovide further embodiments. Aspects of the disclosure can be modified,if necessary, to employ the compositions, functions and concepts of theabove references and application to provide yet further embodiments ofthe disclosure. These and other changes can be made to the disclosure inlight of the detailed description. All such modifications are intendedto be included within the scope of the appended claims.

Specific elements of any of the foregoing embodiments can be combined orsubstituted for elements in other embodiments. Furthermore, whileadvantages associated with certain embodiments of the disclosure havebeen described in the context of these embodiments, other embodimentsmay also exhibit such advantages, and not all embodiments neednecessarily exhibit such advantages to fall within the scope of thedisclosure.

Examples

The following examples illustrate some embodiments and aspects of theinvention. It will be apparent to those skilled in the relevant art thatvarious modifications, additions, substitutions, and the like can beperformed without altering the spirit or scope of the invention, andsuch modifications and variations are encompassed within the scope ofthe invention as defined in the claims which follow. The followingexamples do not in any way limit the invention.

Example 1: Synthesis of Cholesterol-Oxaliplatin Compounds [Formula I]with Carbamate Linkage

Cholesterol-Oxaliplatin complexes comprising Carbamate linkage weresynthesized as follows (FIG. 1):

Part A (FIG. 1A): (Step a):

In a 250 mL round bottom flask, ethylenediamine (about 22.2 mL, 30 eq)was added to about 50 mL of dry DCM (dichloromethane). The reactionflask was cooled to about 0° C. under ice bath. Solid cholesterylchloroformate (about 5.0 g, 11.14 mmol) dissolved in another 50 mL ofdry DCM was added to the reaction flask dropwise with a dropping funnelfor a period of about 30 to about 45 minutes with vigorous stirring. Theresulting solution was stirred at room temperature (25° C.) forovernight (about 8 hours to 12 hours). The solution was thereafter takenin chloroform (about 100 mL) and washed sequentially for about one timeto 3 times with water (3×50 mL) and brine (1×50 mL). The organic layerobtained was dried over anhydrous sodium sulfate, filtered and thesolvent from the filtrate was removed by rotary evaporation. The residueupon column chromatographic purification with 60-120 mesh silica gelusing 1% methanol-chloroform (v/v) as eluent gave 0.4.12 g (78%) of thepure intermediate I [FIG. 1, Part A]. (R_(f)=0.2 using 10%methanol-chloroform v/v, as the TLC developing solvent).

The characterization of intermediate (I) was carried out by proton NMRand the results are as follows: ¹H NMR (500 MHz, CDCl₃) δ 5.30 (s, 1H,—C═CH), 5.05 (s, 1H, —O—CO—NH), 4.42 (s, 1H, —CH—O—), 3.18 (s, 2H,—HN—CH ₂—CH₂—), 2.79 (s, 2H, —O—CO—NH—CH ₂), 2.35- 0.60 (m, 45H,cholesterol backbone).

(Step b):

The intermediate I obtained in step (a) (about 1.0 g, 2.12 mmol) andsuccinic anhydride (about 1.04 g, 10.57 eq) together were dissolved indry DCM (about 20 mL) and were stirred at room temperature (25° C.) fora time-period ranging from about 15 minutes to about 30 minutes.Pyridine (about 3.41 mL, 20 eq) was added dropwise and the reactionmixture was stirred for overnight (about 8 hours to 12 hours). Thereaction mixture was then diluted with about 50 mL of chloroform and waswashed for about three times with 0.1N HCl (3×100 mL) and brine (1×100mL). The organic layer obtained was dried over anhydrous sodium sulfate,filtered and the solvent from the filtrate was removed by rotaryevaporation and thereafter purified by silica gel chromatography toafford about 1.06 g (87%) of intermediate II. (R_(f)=0.2 using 20%Methanol-chloroform v/v, as the TLC developing solvent).

The characterization of intermediate II was carried out by proton NMRand the results are as follows: ¹H NMR (500 MHz, CDCl₃) δ 6.72 (s, 1H,NH), 5.32 (s, 1H, —C═CH), 5.09 (s, 1H, NH), 4.42 (s, 1H, CH—O—),3.35-3.18 (m, 4H, —CO—NH—CH ₂, CO₂H—CH ₂—), 2.65 (s, 2H, —O—CO—NH—CH₂—), 2.48 (s, 2H, —NH—CO—CH ₂—), 2.25-0.62 (m, 43H, cholesterolbackbone). ESIMS m/z=572 [M+1]⁺ for C₃₄H₅₆N₂O₅

(Step c):

In a 50 ml single neck round bottom flask about 0.15 ml (1.27 mmol)monoethylmalonate was taken along with about 185 mg (1.37 mmol) HOBt andabout 263 mg (1.37 mmol) EDCl. About 7 ml dry DCM was added and thereaction mixture was continuously stirred for a time-period of about 20minutes to 30 minutes under N₂ atmosphere. At 0° C., about 500 mg (1.06mmol) of the intermediate I obtained in step (a) [Example 1] dissolvedin about 5 ml dry DCM (20 mL) was added to the reaction mixture. DIPEA(N,N-diisopropylethylamine) was added dropwise until the pH of reactionmixture reached alkaline. The reaction was continuously stirred at roomtemperature (of about 20° C. to 25° C.) for overnight (about 8 hours to12 hours). The reaction mixture was thereafter washed for about threetimes with 0.1 N HCl (1×30 ml), saturated NaHCO₃ (1×50 ml) and brine(1×30 ml). The organic layer obtained was dried over anhydrous Na₂SO₄and evaporation was carried out in rotary evaporator. Columnchromatographic purification (1.5% chloroform-methanol) was performedwhich resulted in yield of about 530 mg (85%) of the intermediate IIIiproduct. (R_(f)=0.6 using 10% Methanol-chloroform v/v, as the TLCdeveloping solvent).

The characterization of the intermediate product IIIi obtained above wascarried out by proton NMR and the results are as follow: ¹H NMR (500MHz, CDCl₃) δ 7.34 (s, 1H, —CH2-NH—CO—), 5.30 (s, 1H, —CH2-CH═C—), 4.90(s, 1H, —OCO—NH—CH2-), 4.42 (s, 1H, —CH—OCO—), 4.14 (m, 2H, —OCH2-CH3),3.48-3.27 (m, 6H), 2.37-0.61 (m, 46H, cholesterol backbone).

(Step d):

In a 50 ml single neck round bottom flask, about 1.03 g (1.75 mmol) ofintermediate obtained in step (c) [Example 1] was taken in THF:H₂O (15ml:5 ml) and the mixture was stirred for about 5 minutes at roomtemperature (about 20° C. to 25° C.). To this reaction mixture, about146 mg (3.50 mmol) of LiOH (Lithium hydroxide) was added and the mixturewas stirred for an additional time-period of about 2-3 hours at roomtemperature (about 25° C.). After completion of the reaction, themixture was diluted with chloroform (about 100 ml) and acidified withabout 50 ml diluted HCl (0.1N). The organic layer obtained was washedwith NaHCO₃ solution (about 50 ml) and dried over anhydrous Na₂SO₄.Column chromatographic purification (4% methanol-chloroform) wasperformed which afforded about 631 mg (64%) of intermediate III.(R_(f)=0.4 using 20% Methanol-chloroform v/v, as the TLC developingsolvent).

(Step e):

In a 50 ml single neck round bottom flask, about 0.12 ml (1.27 mmol)monoethyloxalate was taken along with about 185 mg (1.37 mmol) HOBt andabout 263 mg (1.37 mmol) EDCl. About 7 ml dry DCM was added and thereaction mixture was continuously stirred for a time-period of about 20minutes to about 30 minutes under N₂ atmosphere. At 0° C., about 500 mg(1.06 mmol) of the intermediate I obtained in step (a) [Example 1]dissolved in about 5 ml dry DCM (about 20 mL) was added to the reactionmixture. DIPEA was added dropwise until the pH of reaction mixturereaches alkaline. The reaction mixture was continuously stirred at roomtemperature for overnight. The reaction mixture was washed with 0.1N HCl(1×30 ml), saturated NaHCO₃ (1×50 ml) and brine (1×30 ml). The organiclayer obtained was dried over anhydrous Na₂SO₄ and evaporated in rotaryevaporator. Column chromatographic purification (1.5%methanol-chloroform) was performed which yielded about 570 mg (94%) ofintermediate product IVi. (R_(f)=0.6 using 10% Methanol-chloroform v/v,as the TLC developing solvent).

The characterization of the intermediate product IVi obtained above wascarried out by proton NMR and the results are as follows: ¹H NMR (500MHz, CDCl₃) δ 7.40 (s, 1H, —CO—NH—CH2-), 5.28 (s, 1H, —CH═C—), 4.30 (q,J=7.1 Hz, 2H, —O—CH2-CH3), 3.52 (d, J=3.7 Hz, 2H, —O—CH2-CH2-), 3.46 (d,J=4.2 Hz, 2H, —NH—CH2-CH2-), 3.11 (t, J=11.0 Hz, 1H, —O—CH—CH2-),2.33-0.55 (m, 47H, cholesterol back bone).

(Step d′):

In a 50 ml single neck round bottom flask, about 500 mg (0.87 mmol)intermediate obtained in step (e) [Example 1] was taken in THF:H₂O (15ml:5 ml) and stirred for about 5 minutes at room temperature. To thisreaction mixture, about 75 mg (1.75 mmol) of LiOH was added and themixture was stirred for an additional time-period of about 2 hours atroom temperature. After completion of the reaction, the mixture wasdiluted with chloroform (about 50 ml) and acidified with about 50 mldiluted HCl (0.1N). The organic layer was washed with NaHCO₃ solution(50 ml) and dried over anhydrous Na₂SO₄. Column chromatographicpurification (4% methanol-chloroform) was performed which afforded about180 mg (38%) of intermediate IV. (R_(f)=0.2 using 10%Methanol-chloroform v/v, as the TLC developing solvent).

Part B (FIG. 1B): (Step f):

Dichloro(1,2-diamino-cyclohexane)platinum (II) (about 300 mg, 0.79 mmol)was partially dissolved in about 40.0 mL of H₂O. To the solution, silvernitrate (about 340 mg, 1.58 mmol) was added and the resulting reactionmixture was stirred at room temperature for a time-period of about 24hours. When the mixture appeared milky white, silver chloride wasremoved by centrifuging at about 12000 rpm for about 30 minutes.Finally, the aquated oxaliplatin V was obtained by filtration through0.2 μM filter.

Part C (FIG. 1C): (Step g): Synthesis of Compound 1:

Intermediate II (about 407 mg, 0.71 mmol) obtained in step b (Example 1,PART A) was dissolved in about 1.5 mL DMF. To the solution, about 40.0mL of aquated oxaliplatin V (0.09 mmol) obtained in step f (Example 1,PART B) was added and the reaction mixture was stirred for about 24hours. Lyophilization of the reaction mixture yieldedcholesterol-oxaliplatin compound Compound 1.

The characterization results (proton NMR and MALDI-TOF MS) of Compound 1are as follow: ¹H NMR (500 MHz, CDCl₃) δ 6.65 (s, 1H, NH), 5.30 (s, 1H,—C═CH), 5.01 (s, 1H, NH), 4.42 (s, 1H, CH—O—), 3.42 (s, 2H, Pt—NH₂—CH—),3.38-3.18 (m, 4H, —CO—NH—CH ₂, CO₂H—CH ₂—), 2.65 (s, 2H, —O—CO—NH—CH₂—), 2.48 (s, 2H, —NH—CO—CH ₂—), 2.30-0.58 (m, 55H, cholesterol backboneand amino cyclohexane). MALDI-TOF MS=880.4784 [M]⁺ for C₄₀H₆₉N₄O₅Pt.

Step (g′): Synthesis of Compound 2:

Intermediate III (about 58 mg, 0.11 mmol) obtained in step d (Example 1,PART A) was dissolved in about 1.5 mL DMF. To the solution, about 8.0 mLof aquated oxaliplatin V (0.11 mmol) obtained in step f (Example 1, PARTB) was added and the reaction mixture was stirred for about 24 hours.Lyophilization of the reaction mixture afforded cholesterol-oxaliplatincompound Compound 2.

The characterization results (proton NMR and MALDI-TOF MS) of Compound 2are as follows: ¹H NMR (500 MHz, CDCl₃) δ 7.41 (s, H, —CO—NH—CH2-), 5.40(s, 1H, —CH2-CH═C—), 5.07 (s, 1H, —OCO—NH—CH2-), -), 4.49 (m, 1H,—CH—OCO—), 3.48-3.27 (m, 6H), 2.88 (s, 1H, —CH—NH2), 2.81 (s, 1H,—CH—NH2), 2.30-0.51 (m, 46H, cholesterol backbone and aminocyclohexane). MALDI-TOF MS=886.5048 [M]⁺ for C₃₉H₆₇N₄O₅Pt.

Step (g″): Synthesis of Compound 3:

Intermediate IV (about 57 mg, 0.11 mmol) obtained in step d′ (Example 1,PART A) was dissolved in about 1.5 mL DMF. To the solution, about 8.0 mLof aquated oxaliplatin V (0.11 mmol) obtained in step f (Example 1, PARTB) was added and the reaction mixture was stirred for about 24 hours.Lyophilization of the reaction mixture afforded cholesterol-oxaliplatincompound Compound 3.

The characterization results (proton NMR and MALDI-TOF MS) of Compound 3are as follows: ¹H NMR (500 MHz, CDCl₃) δ 7.84 (s, 1H, —O—CO—NH—CH2-),5.31 (s, 1H, —CH═C—), 4.89 (s, 1H, —CO—NH—CH2-), 4.44 (s, 1H,—O—CH—CH2-), 3.48-3.40 (m, 2H, —CO—NH—CH2-), 3.37-3.28 (m, 2H,—O—CO—NH—CH2-), 2.91 (s, 1H, NH2-CH—CH2-), 2.83 (s, 1H, NH2-CH—CH2-),2.33-0.56 (m, 55H, cholesterol back bone, and cyclohexane ring protons).MALDI-TOF MS=853.4823 [M]⁺ for C₃₈H₆₅N₄O₅Pt

Example 2: Synthesis of Cholesterol-Oxaliplatin Compounds [Formula I]with Ether Linkage

Cholesterol-Oxaliplatin complexes comprising ether linkage weresynthesized as follows (FIG. 2):

PART A (FIG. 2A): (Steps a-e):

Synthesis of amine intermediate (II): Steps a to e was carried out asdescribed in Example 3 (Steps 1 to 5; Synthesis of Compound 25).

(Step f):

To a 100 mL single round bottom flask, intermediate II obtained aftersteps (a-e) [Example 2, PART A] (amine 500 mg, 1.164 mmol) was taken inDCM (about 10 mL) under N₂ atmosphere and stirred for a time-periodranging from about five minutes to about ten minutes at roomtemperature. The reaction mixture was cooled to about 0° C. and succinicanhydride (about 570 mg, 5.82 mmol) followed by pyridine (about 1.88 ml,23.3 mmol) was added and the reaction mixture was again allowed to stirfor about 24 hours at room temperature.

After completion of the stirring process (checked by TLC), the reactionmixture was diluted with CH₂Cl₂ (about 20 mL) and washed with 0.1 N HCl(about 500 mL, to remove pyridine completely) followed by drying overanhydrous Na₂SO₄. The organic layer was concentrated under vacuo andpurified by silica gel chromatography which afforded the requiredintermediate III in 95% yield (about 585 mg).

The characterization of intermediate III was carried out by proton NMRand the results are as follows: ¹H NMR (500 MHz, CDCl₃) δ 6.18 (s, 1H,—CO—NH—CH2-), 5.28 (s, 1H, —CH═C—), 3.49 (d, J=4.4 Hz, 2H, —O—CH2-CH2-),3.38 (d, J=4.4 Hz, 2H, —NH—CH2-CH2-), 3.11 (t, J=11.1 Hz, 1H,—O—CH—CH1-), 2.63 (t, J=6.4 Hz, 2H, —NH—CO—CH2-), 2.47 (t, J=6.4 Hz, 2H,HOOC—CH2-), 2.31-0.55 (m, 43H, cholesterol back bone).

(Step g):

In a 50 ml single neck round bottom flask, about 0.07 ml (0.64 mmol)monoethylmalonate was taken with about 74 mg (0.64 mmol) HOBt and about134 mg (0.69 mmol) EDCl. About 7 ml dry DCM was added and reactionmixture was continuously stirred for a time-period of about 20 minutesto 30 minutes under N₂ atmosphere. At about 0° C., about 250 mg (0.58mmol) of the intermediate II obtained after steps (a-e) [Example 2, PARTA] dissolved in about 5 ml dry DCM (20 mL) was added to the reactionmixture. DIPEA was added dropwise until the pH of reaction mixtureturned alkaline. The reaction mixture was continuously stirred at aboutroom temperature for overnight and washed for about 3-4 times with 0.1NHCl (1×30 ml), saturated NaHCO₃ (1×50 ml) and brine (1×30 ml)successively. The organic layer was dried over anhydrous Na₂SO₄ andevaporation was carried out in rotary evaporator. Column chromatographicpurification (1.5% methanol-chloroform) was performed which yieldedabout 290 mg (92% pure) of intermediate IVi compound (R_(f)=0.5 using 5%Methanol-chloroform v/v, as the TLC developing solvent).

The characterization of the intermediate product IVi obtained above wascarried out by proton NMR and the results are as follows: ¹H NMR (500MHz, CDCl₃) δ 7.27 (s, 1H, —CO—NH—CH2-), 5.28 (s, 1H, —CH═C—), 4.13 (q,J=7.1 Hz, 2H, —O—CH2-CH3), 3.49 (d, J=4.0 Hz, 2H, —O—CH2-CH2-), 3.40 (d,J=4.8 Hz, 2H, —NH—CH2-CH2-), 3.25 (s, 2H, —CO—CH2-CO—), 3.10 (t, J=10.9Hz, 1H, —O—CH—CH2-), 2.34-0.55 (m, 46H, cholesterol back bone).

(Step h):

In a 25 ml single neck round bottom flask, about 250 mg (0.46 mmol)intermediate IVi obtained in step (g) [Example 2] was taken in THF:H₂O(9 ml:3 ml) and the mixture was stirred for about 5 minutes at roomtemperature. To this reaction mixture, about 58 mg (1.38 mmol) LiOH wasadded and the mixture was stirred for an additional time-period of about3 hours at room temperature (about 20° C. to 25° C.). After completionof the reaction, the mixture was diluted with chloroform (about 50 ml)and acidified with about 10 ml diluted HCl (0.1N). The organic layer waswashed with NaHCO₃ solution (about 20 ml) and dried over anhydrousNa₂SO₄. Column chromatographic purification (4% methanol-chloroform) wasperformed which afforded about 228 mg (96%) of intermediate IV.(R_(f)=0.4 using 20% Methanol-chloroform v/v, as the TLC developingsolvent).

(Step i):

In a 50 ml single neck round bottom flask, about 0.06 ml (10.64 mmol)monoethyloxalate was taken along with about 74 mg (0.64 mmol) HOBt andabout 145 mg (0.76 mmol) EDCl. About 7 ml dry DCM was added and thereaction mixture was continuously stirred for a time-period of about 20minutes to about 30 minutes under N₂ atmosphere. At 0° C., about 250 mg(0.58 mmol) of the intermediate II obtained after steps (a-e) dissolvedin about 5 ml dry DCM (20 mL) was added to the reaction mixture. DIPEAwas added dropwise until the pH of the reaction mixture turned alkaline.The reaction mixture was continuously stirred at room temperature forovernight followed by washing with 0.1N HCl (1×30 ml), saturated NaHCO₃(1×50 ml) and brine (1×30 ml) successively. The organic layer was driedover anhydrous Na₂SO₄ and evaporated in rotary evaporator. Columnchromatographic purification (1.5% methanol-chloroform) was performedwhich yielded about 128 mg (41%) of intermediate VIi compound.(R_(f)=0.5 using 10% Methanol-chloroform v/v, as the TLC developingsolvent).

The characterization of the intermediate product VIi obtained above wascarried out by proton NMR and the results are as follows: ¹H NMR (500MHz, CDCl₃) δ 7.40 (s, 1H, —CO—NH—CH2-), 5.28 (s, 1H, —CH═C—), 4.30 (q,J=7.1 Hz, 2H, —O—CH2-CH3), 3.52 (d, J=3.7 Hz, 2H, —O-CH2-CH2-), 3.46 (d,J=4.2 Hz, 2H, —NH—CH2-CH2-), 3.11 (t, J=11.0 Hz, 1H, —O—CH—CH2-),2.33-0.55 (m, 47H, cholesterol back bone).

Step (h′):

In a 25 ml single neck round bottom flask, about 128 mg (0.22 mmol)intermediate as obtained in step (i) [Example 2] above was taken inTHF:H₂O (3 ml:1 ml) and stirred for about 5 minutes at room temperature.To this reaction mixture, about 18 mg (0.45 mmol) LiOH was added and thereaction mixture was stirred for an additional time-period of about 2hours at room temperature. After completion of the reaction, thereaction mixture was diluted with chloroform (about 15 ml) followed byacidification with about 10 ml diluted HCl (0.1N). The organic layer waswashed with NaHCO₃ solution (about 20 ml) and dried over anhydrousNa₂SO₄. Column chromatographic purification (4% methanol-chloroform) wasperformed which afforded about 79 mg (66% pure) of intermediate V.(R_(f)=0.2 using 10% Methanol-chloroform v/v, as the TLC developingsolvent).

Part B (FIG. 2B): Step (i):

Dichloro (1,2-diamino-cyclohexane) platinum (II) (about 300 mg, 0.79mmol was partially dissolved in about 40.0 mL of H₂O. Silver nitrate(about 340 mg, 1.58 mmol) was added to the same and the resultingreaction mixture is stirred at room temperature for about 24 hours.After the appearance of milky white, silver chloride was removed bycentrifuging at around 12000 rpm for about 30 minutes. Finally, theaquated oxaliplatin VI was obtained by filtration through 0.2 μM filter.

Part C (FIG. 2C): Step (k): Synthesis of Compound 4:

Intermediate III (about 69 mg, 0.13 mmol) obtained in step (f) (Example2, PART A) was dissolved in about 1.5 mL DMF. Thereafter, about 10 mL ofaquated oxaliplatin VI (0.13 mmol) as obtained in step (j) (Example 2,PART B) was added and the reaction mixture was stirred for about 24hours. Lyophilization of the reaction mixture affordscholesterol-oxaliplatin compound Compound 4.

The characterization results (proton NMR and MALDI-TOF MS) of Compound 4are as follows: ¹H NMR (500 MHz, CDCl₃) δ 6.12 (s, 1H, —CO—NH—CH2-),5.30 (d, J=4.9 Hz, 1H, —CH═C—), 3.50 (t, J=4.8 Hz, 2H, —O—CH2-CH2-),3.43-3.35 (m, 2H, —NH—CH2-CH2-), 3.17-3.05 (m, 1H, —O—CH—CH2-), 2.91 (s,1H, NH2-CH—), 2.84 (s, 1H, NH2-CH—), 2.65 (dd, J=7.6, 5.1 Hz, 2H,—O—CO—CH2-CH2), 2.56-2.46 (m, 2H, —NH—CO—CH2-), 2.30-0.56 (m, 55H,cholesterol back bone, and cyclohexane ring protons). MALDI-TOFMS=837.5227 [M]⁺ for C₃₉H₆₈N₃O₄Pt.

Step (k′): Synthesis of Compound 5:

Intermediate IV (27 mg, 0.05 mmol) obtained in step h (Example 2, PARTA) was dissolved in about 1.5 mL DMF. Thereafter, about 10 mL of aquatedoxaliplatin VI (0.05 mmol) obtained in step j (Example 2, Part B) wasadded and the reaction mixture was stirred for about 24 hours.Lyophilization of the reaction mixture afforded cholesterol-oxaliplatincompound Compound 5.

The characterization results (proton NMR and MALDI-TOF MS) of Compound 5are as follows: ¹H NMR (500 MHz, CDCl₃) δ 6.52 (s, 1H, —CO—NH—CH2-),5.28 (d, J=5.0 Hz, 1H, —CH═C—), 3.51 (t, J=4.5 Hz, 2H, —O—CH2-CH2-),3.41 (m, 2H, —NH—CH2-CH2-), 3.27 (s, 2H, —CO—CH2-CO—), 3.17-3.02 (m, 1H,—O—CH—CH2-), 2.89 (s, 1H, NH2-CH—), 2.82 (s, 1H, NH2-CH—), 2.30-0.56 (m,55H, cholesterol back bone, and cyclohexane ring protons). MALDI-TOFMS=823.5242 [M]⁺ for C₃₈H₆₆N₃O₄Pt

Step (k″): Synthesis of Compound 6:

Intermediate V (26 mg, 0.05 mmol) obtained in step h′ (Example 2, PARTA) was dissolved in about 1.5 mL DMF. Thereafter, about 5 mL of aquatedoxaliplatin VI (0.05 mmol) obtained in step e (Example 2, PART B) wasadded and the reaction mixture was stirred for about 24 hours.Lyophilization of the reaction mixture afforded cholesterol-oxaliplatincompound Compound 6.

The characterization results (proton NMR and MALDI-TOF MS) of Compound 6are as follows: ¹H NMR (500 MHz, CDCl₃) δ 7.96 (s, 1H, —CO—NH—CH2-),5.28 (s, 1H, —CH═C—), 3.61-3.42 (m, 4H, —OCH2-CH2-, —NH—CH2-CH2),3.18-3.02 (m, 1H, —OCH—CH2-), 2.90 (s, 1H, NH2-CH—), 2.82 (s, 1H,NH2-CH—), 2.31-0.56 (m, 55H, cholesterol back bone, and cyclohexane ringprotons). MALDI-TOF MS=809.5258 [M]⁺ for C₃₇H₆₄N₃O₄Pt

Similar to the synthetic procedures as described above, Compound 7-21were prepared by employing necessary carboxylic acids, linker molecules,lipid and platinum moieties.

Example 3: Synthesis of Compounds of Formula II Synthesis of Compound 25

Step 1:

To an ice cooled solution of cholesterol 1.01 (about 10 g, 0.026 mol) inCH₂Cl₂ (about 45 mL), pyridine (about 15 mL) is added and stirred forabout 15 minutes. To this solution, p-toluene sulphonyl chloride (about9.8 g, 0.052 mol) is added and stirred for about 6 h at about 0° C. andthereafter, TLC is checked. After completion, the reaction mixture isdiluted with CHCl₃ (about 20 mL) and washed with about 1N HCl (3×50 mL)and brine (about 20 mL) successively. The organic layer is dried overanhydrous Na₂SO₄ and concentrated under vacuum to afford intermediate1.02 and the said intermediate is directly taken for the next reactionwithout further purification.

Step 2:

To the solution of tosylated cholesterol 1.02 (about 10 g, 0.018 mol) indioxane (about 45 mL), ethylene glycol (about 15 mL) is added andrefluxed for about 4 h. The TLC is checked. After completion, thereaction mixture is extracted with ethyl acetate and washed with water(about 3×50 mL) and brine (about 20 mL) successively. The organic layeris dried over anhydrous Na₂SO₄ and concentrated under vacuum and columnpurified to afford intermediate 1.03.

Step 3:

To an ice cooled solution of cholesteryl ethylene glycol 1.03 (about6.95 g, 16.13 mmol) in dichloro methane (about 15 ml) pyridine (about 13mL) is added under nitrogen atmosphere and stirred for about 15 minutes.To this solution, p-toluene sulphonyl chloride (about 3.7 g, 19.35 mmol)is added and stirred for about 5 h at about 0° C. and TLC is checked.After completion, the reaction mixture is diluted with CHCl₃ (about 20mL) and washed with about 1N HCl (3×50 mL) and brine (about 20 mL)successively. The organic layer is dried over anhydrous Na₂SO₄ andconcentrated under vacuum and purified by silica gel chromatography toobtain intermediate 1.04.

Step 4:

To a 50 mL round bottomed flask, compound 1.04 (about 6 g, 10.26 mmol)is taken in DMF (about 20 ml) under nitrogen atmosphere and is stirredfor about 30 minutes to get a clear solution (warm if necessary). Tothis solution, sodium azide (about 3.4 g, 51.33 mmol) is added andstirred for about 18 h at room temperature and TLC is checked. Aftercompletion, the reaction mixture is concentrated under vacuum to removeTHF and is purified by flash chromatography to obtain intermediate 1.05.

Step 5:

To a solution of azide 1.05 (about 3 g, 7.6 mmol) in dry DMF (about 15ml), TPP (about 1.5 g, 15.2 mmol) is added under nitrogen atmosphere.The reaction is stirred for about 6 h at room temperature and about 2 mLof water is added to the reaction mixture. The reaction mixture isstirred for additional time-period of about 6 h and TLC is checked.After completion, the reaction mixture is concentrated under reducedpressure and is purified by silica gel chromatography utilizingmethanol/chloroform as eluent to achieve amine intermediate 1.06.

Step 6:

To an ice cool solution of amine 1.06 (about 300 mg, 0.698 mmol) in THF(about 5 mL), NaH (about 120 mg, 2.094 mmol) is added by pinch over aperiod of about 10 minutes. The resulting solution is stirred for about20 minutes and ethyl bromo acetate is added and stirred for atime-period of about 6 h at room temperature. After completion, thereaction mixture is cooled to about 0° C. and quenched with water andthe compound is extracted with ethyl acetate (about 2×20 mL). Theorganic layer is dried over anhydrous Na₂SO₄, concentrated and purifiedby silica gel chromatography to obtain diester intermediate 1.07 inabout 52% yield.

Step 7:

To a 50 mL single neck round bottom flask, diester compound 1.07 (about218 mg, 0.363 mmol) is taken in THF/water (about 4 mL, at a ratio ofabout 3:1) and cooled to about 0° C. To this cooled solution, LiOH(about 34 mg, 1.45 mmol) is added and stirred at room temperature for atime-period of another 6 h. After completion, the reaction mixture isconcentrated under reduced pressure to remove THF and the aqueous layeris washed with ethyl acetate. The aqueous layer is lyophilized to getsolid di-lithium salt of 1.08 with a quantitative yield.

Step 8:

Synthesis of DACH-Pt(H₂O)₂: To a 50 mL single neck round bottom flaskdichloro (1,2-diamino-cyclohexane) platinum 1.09 (about 200 mg, 0.526mmol) is taken in about 20.0 mL of H₂O. To this suspension, silvernitrate (about 178.7 mg, 1.052 mmol) is added and the reaction mixtureis stirred at room temperature for about 24 h. The milky white solutionis centrifuged and the solution is filtered through 0.22 μM syringefilter to obtain aquated DACH-Pt 1.10 in quantitative yield (about 10mg/mL).

Step 9:

To a 100 mL single neck round bottom flask intermediate 1.08 (about 202mg, 0.363 mmol) is taken in about 1.0 mL water. To this solution,DACHPt(H₂O)₂ (about 13.8 mL) obtained in the previous step is added andstirred for another 12 h. The solid residue is filtered and washed withwater (about 20 mL). The white solid residue is lyophilized anddissolved in excess methanol, filtered and concentrated under reducedpressure to afford cholesterol-oxaliplatin amphiphile Compound 25 inabout 85% yield.

Synthesis of Compound 26

Step 1:

To an ice cooled solution of ethylene diamine (about 22.2 mL) in CH₂Cl₂(about 40 mL), a solution of compound 1.11 (about 5 g) in CH₂Cl₂ (about50 mL) is added dropwise over a period of about 45 min and the reactionmixture is stirred at the same temperature for about 1 h and is furtherallowed to stir at room temperature for an additional time-period ofabout 20 h. After completion (checked by TLC), the reaction mixture isquenched with water and extracted with dichloro methane (about 4×50 mL),the combined organic layer is dried over anhydrous Na₂SO₄ andconcentrated under vacuum. The residue is purified by columnchromatography utilizing methanol-chloroform as eluent to obtainintermediate 1.12.

Step 2:

To a 50 mL single neck round bottom flask amine 1.12 (about 300 mg 0.634mmol) is taken in THF (about 5 mL) under nitrogen atmosphere. Thereaction mixture is cooled to about 0° C. under ice bath and NaH (about130 mg, 3.17 mmol) is added by pinch over a period of about 10 minutes.The resulting solution is stirred for about 20 minutes and ethyl bromoacetate is added. The reaction mixture is stirred for about 2 h at roomtemperature and TLC is checked. After completion, the reaction mixtureis cooled to about 0° C. and quenched with cold water (about 5 mL),extracted with ethyl acetate (about 2×20 mL), dried over anhydrousNa₂SO₄ and thereafter concentrated. The residue is purified by columnchromatography utilizing methanol-chloroform as eluent to obtainintermediate 1.13.

Step 3:

To a 50 mL single neck round bottom flask, diester 1.13 (about 1.7 g,2.63 mmol) is taken in THF/water (about 3:1) (about 16 mL). The reactionmixture is cooled to about 0° C. under ice bath and LiOH (about 130 mg,5.27 mmol) is added to the reaction mixture. The resulting solution isstirred for about 6 h at room temperature and TLC is checked. Aftercompletion, the reaction mixture is concentrated under reduced pressureto remove THF and diluted with water (about 5 mL). The water layer iswashed with ethyl acetate and CH₂Cl₂ successively and lyophilized toobtain intermediate 1.14 in quantitative yield.

Step 4:

To a 100 mL single neck round bottom flask, intermediate 1.14 is takenin about 1.0 mL water. To this solution, DACHPt(H₂O)₂ is added andstirred for a time-period of another 12 h. The solid residue obtained isfiltered, washed with water and lyophilized. The residue is dissolved inexcess methanol, filtered and concentrated under reduced pressure toafford cholesterol-oxaliplatin amphiphile Compound 26.

Synthesis of Compound 27

Step 1:

To a 50 mL single neck round bottom flask, BocHNCH₂COOH (about 370 mg,2.08 mmol) is taken in CH₂Cl₂ (about 10 mL) under nitrogen atmosphere.Solid EDCl (about 400 mg, 2.08 mmol) and HOBT (about 285 mg, 2.08 mmol)are added successively to the reaction mixture. DIPEA is added to makethe solution alkaline and the reaction mixture is stirred for another 20minutes. To this activated acid solution, amine 1.06 (about 450 mg, 1.04mmol) is added and the mixture is stirred at room temperature for about12 hand TLC is checked. Ater completion, the reaction mixture isquenched with water, extracted with chloroform, dried over anhydrousNa₂SO₄ and thereafter concentrated. The residue is purified by silicagel chromatography utilizing methanol-chloroform as eluent to obtainintermediate 1.15.

Step 2:

To a 50 mL single neck round bottom flask, Boc protected amine 1.15(about 600 mg, 0.99 mmol) is taken in CH₂Cl₂ and the flask is cooled toabout 0° C. To this solution, TFA is added and the mixture is stirredfor about 3 hours at the same temperature. After completion, thereaction mixture is concentrated under rotary evaporator and the crudeproduct 1.16 is utilized for the next reaction without furtherpurification.

Step 3:

To a 50 mL single neck round bottom flask, crude amine 1.16 (about 400mg, 0.821 mmol) is taken in THF (about 10 mL) under nitrogen atmosphere.The solution is cooled to about 0° C. under ice bath and solid NaH(about 160 mg, 4.10 mmol) is added pinch wise over a period of about 10minutes. The resulting solution is stirred for an additional 20 minutesand ethyl bromo acetate is added. After completion, the reaction mixtureis cooled to about 0° C. and quenched with water, extracted with ethylacetate, dried over anhydrous Na₂SO₄ and thereafter concentrated undervacuum. The residue is purified by silica gel chromatography utilizingmethanol-chloroform as eluent to obtain intermediate 1.17.

Step 4:

To a 50 mL single neck round bottom flask, diester 1.17 (about 200 mg,0.303 mmol) is taken in THF/water (about 3:1) (about 4 mL) at about 0°C. Solid LiOH (about 15 mg, 0.606 mmol) is added to the reaction mixtureand stirred for about 6 hours at room temperature. After completion, thereaction mixture is concentrated and diluted with water (about 4 mL).The aqueous layer is washed with ethyl acetate and dichloro methanesuccessively and is lyophilized to obtain solid powder of acid salt 1.18in quantitative yield.

Step 5:

To a 100 mL single neck round bottom flask, intermediate 1.18 is takenin about 1.0 mL water. To this solution, DACHPt(H₂O)₂ is added andstirred for another 12 hours. The solid residue is filtered and washedwith water and thereafter lyophilized. The residue is dissolved inexcess methanol, filtered and concentrated under reduced pressure toafford cholesterol-oxaliplatin amphiphile Compound 27.

Synthesis of Compound 28

Step 1:

To the solution of intermediate 1.02 (about 6 g, 0.011 mol) in dioxane(about 30 mL) is added diethylene glycol (about 20 mL) and allowed toreflux for about 4 hours. After completion, the reaction mixture isquenched with water (about 20 mL) and extracted with ethyl acetate. Theorganic layer is washed with water (about 3×50 mL) and brine (about 20mL) successively and dried over anhydrous Na₂SO₄. The combined organiclayer is concentrated under reduced pressure and the residue is purifiedby silica gel chromatography utilizing methanol-chloroform as eluent toobtain intermediate 1.19.

Step 2:

To an ice cooled solution of cholesteryl alcohol 1.19 (about 5 g, 10.54mmol) and p-toluene sulphonyl chloride (about 4 g, 21.09 mmol) in DCM(about 25 ml) under nitrogen atmosphere, pyridine (about 13 mL) isadded. The solution is stirred for about 5 hours at about 0° C. and TLCis checked. After completion, the solution is diluted with CHCl₃ (about20 mL) and washed with about 10% copper (II) sulphate solution (about3×50 mL) and brine (about 20 mL) successively. The organic layer isdried over anhydrous Na₂SO₄ and concentrated under reduced pressure. Theresidue is purified by silica gel chromatography utilizing ethylacetate/hexane as eluent to obtain intermediate 1.20.

Step 3:

To a 50 mL round bottom flask tosyl compound 1.20 (about 3 g, 4.76 mmol)is taken in DMF (about 15 ml) under nitrogen atmosphere and stirred forabout 30 minutes to get a clear solution (warm if necessary). To thissolution, solid sodium azide (about 1.55 g, 23.84 mmol) is added andstirred for about 18 h at room temperature and TLC is checked. Aftercompletion, the reaction mixture is diluted with water (about 50 mL),extracted with ethyl acetate (about 3×20 mL). The organic layer iswashed with water (about 2×20 mL) and brine (about 20 mL) successively.The combined organic layer is dried over anhydrous Na₂SO₄, concentratedunder reduced pressure and purified by flash chromatography to obtainintermediate 1.21.

Step 4:

To a solution of azide 1.21 (about 1 g, 2.01 mmol) in dry DMF (about 10ml) triphenyl phosphene (TPP) (about 1.04 g, 4.02 mmol) is added undernitrogen atmosphere. The reaction mixture is stirred for about 6 hoursat room temperature and water (about 1 mL) is added to it. The resultingsolution is stirred for an additional 6 hours at the same temperatureand TLC is checked. After completion, organic solvent is removed undervacuum and the residue is purified by silica gel chromatographyutilizing methanol/chloroform as eluent to obtain amine intermediate1.22.

Step 5:

To an ice cool solution of amine 1.22 (about 800 mg, 1.68 mmol) in THF(about 10 mL), NaH (about 200 mg, 5.02 mmol) is added under nitrogenatmosphere over a period of about 10 minutes. The resulting solution isstirred for about 20 minutes and ethyl bromo acetate (about 0.78 mL,6.72 mmol) is added and stirred for another 6 hours at room temperature.After completion, the reaction mixture is cooled to about 0° C. andquenched with water and extracted with ethyl acetate (about 2×20 mL).The combined organic layer is dried over anhydrous Na₂SO₄, concentratedand purified by silica gel chromatography to obtain diester intermediate1.23.

Step 6:

To a 50 mL single neck round bottom flask diester 1.23 (about 220 mg,0.340 mmol) is taken in THF/water (about 3:1) (about 4 mL) at about 0°C. Solid LiOH (about 20 mg, 0.640 mmol) is added to the reaction mixtureand stirred for about 6 hours at room temperature. After completion, thereaction mixture is concentrated and diluted with water (about 4 mL).The aqueous layer is washed with ethyl acetate and dichloro methanesuccessively and lyophilized to obtain solid powder of acid salt 1.24 inquantitative yield.

Step 7:

To a 50 mL single neck round bottom flask, dilithium salt 1.24 is takenin water (about 1.0 mL). To this solution, DACHPt(H₂O)₂ is added andstirred for a time-period of another 12 hours and thereafter filtered.The solution is lyophilized to afford cholesterol-oxaliplatin amphiphileCompound 28.

Synthesis of Compound 29

Step 1:

To an ice cool solution of amine 1.06 (about 300 mg, 0.673 mmol) in THF(about 10 mL), NaH (about 108 mg, 2.692 mmol) is added over a period ofabout 10 minutes under nitrogen atmosphere. The resulting solution isstirred for about 20 minutes and ethyl bromo acetate (about 0.1 mL,0.874 mmol) is added. The resulting solution is stirred for another 6hours at room temperature and TLC is checked. After completion, thereaction mixture is cooled to about 0° C. and quenched with water andextracted with ethyl acetate (about 2×15 mL). The combined organic layeris dried over anhydrous Na₂SO₄, concentrated and purified by silica gelchromatography to obtain diester intermediate 1.25.

Step 2:

To a 50 mL single neck round bottom flask, diester 1.25 (about 200 mg,0.388 mmol) is taken in THF/water (about 3:1) (about 4 mL) at about 0°C. Solid LiOH (about 38 mg, 1.55 mmol) is added to the reaction mixtureand stirred for about 6 hours at room temperature. After completion, thereaction mixture is concentrated and diluted with water (about 4 mL).The aqueous layer is acidified with NaHSO₄ solution, extracted withdichloro methane (about 3×10 mL) and further concentrated to obtain asolid powder of acid 1.26 in good yield.

Step 3:

To a 50 mL single neck round bottom flask acid 1.26 (about 70 mg, 0.143mmol) is taken in about 1.0 mL DMF. To this solution, DACH-Pt(H₂O)₂ isadded and stirred for a time-period of another 12 hours. The reactionmixture is lyophilized to afford cholesterol-oxaliplatin amphiphileCompound 29.

Synthesis of Compounds 38, 39, 40, 41 and 42

The synthetic methods for compounds 38, 39, 40, 41 and 42 are similar tothe method of synthesis of compound 25. The lipid moiety varies in thesaid compounds. The different lipid moieties (R) present in compounds38-42 are as follows:

The synthetic routes of said compounds 38-42 are provided in FIG. 4.

Modified Synthesis of Compound 25

Step 1:

To an ice cooled solution of cholesterol 1.01 (about 10 g, 0.026 mol) inCH₂Cl₂ (about 45 mL), pyridine (about 15 mL) was added and stirred forabout 15 minutes. To this solution, p-toluene sulphonyl chloride (about9.8 g, 0.052 mol) was added and stirred for about 6 h at 0° C. andthereafter, TLC was checked. After completion, the reaction mixture wasdiluted with CHCl₃ (about 20 mL) and washed with about 1N HCl (3×50 mL)and brine (about 20 mL) successively. The organic layer was dried overanhydrous Na₂SO₄ and concentrated under vacuum to afford intermediate1.02 and the said intermediate was directly taken for the next reactionwithout further purification.

Step 2:

To the solution of tosylated cholesterol 1.02 (about 10 g, 0.018 mol) indioxane (about 45 mL), ethylene glycol (about 15 mL) was added andrefluxed for about 4 h. The TLC was checked. After completion, thereaction mixture was extracted with ethyl acetate and washed with water(about 3×50 mL) and brine (about 20 mL) successively. The organic layerwas dried over anhydrous Na₂SO₄ and concentrated under vacuum and columnpurified to afford intermediate 1.03.

Step 3:

To an ice cooled solution of cholesteryl ethylene glycol 1.03 (about6.95 g, 16.13 mmol) in dichloro methane (about 15 ml) pyridine (about 13mL) was added under nitrogen atmosphere and stirred for about 15minutes. To this solution, p-toluene sulphonyl chloride (about 3.7 g,19.35 mmol) was added and stirred for about 5 h at about 0° C. and TLCwas checked. After completion, the reaction mixture was diluted withCHCl₃ (about 20 mL) and washed with about 1N HCl (3×50 mL) and brine(about 20 mL) successively. The organic layer was dried over anhydrousNa₂SO₄ and concentrated under vacuum and purified by silica gelchromatography to obtain intermediate 1.04.

Step 4:

To a 50 mL round bottomed flask, compound 1.04 (about 6 g, 10.26 mmol)was taken in DMF (about 20 ml) under nitrogen atmosphere and was stirredfor about 30 minutes to get a clear solution (warm if necessary). Tothis solution, sodium azide (about 3.4 g, 51.33 mmol) was added andstirred for about 18 h at room temperature and TLC was checked. Aftercompletion, the reaction mixture was concentrated under vacuum to removeTHF and was purified by flash chromatography to obtain intermediate1.05.

Step 5:

To a solution of azide 1.05 (about 3 g, 7.6 mmol) in dry DMF (about 15ml), TPP (about 1.5 g, 15.2 mmol) was added under nitrogen atmosphere.The reaction was stirred for about 6 h at room temperature and about 2mL of water was added to the reaction mixture. The reaction mixture wasstirred for additional time-period of about 6 h and TLC was checked.After completion, the reaction mixture was concentrated under reducedpressure and was purified by silica gel chromatography utilizingmethanol/chloroform as mobile phase to achieve amine intermediate 1.06.

Step 6:

To an ice cool solution of amine 1.06 (about 300 mg, 0.698 mmol) in THF(about 5 mL), NaH (about 120 mg, 2.094 mmol) was added by pinch over aperiod of about 10 minutes. The resulting solution was stirred for about20 minutes and ethyl bromo acetate was added and stirred for atime-period of about 6 h at room temperature. After completion, thereaction mixture was cooled to about 0° C. and quenched with water andthe compound was extracted with ethyl acetate (about 2×20 mL). Theorganic layer was dried over anhydrous Na₂SO₄, concentrated and purifiedby silica gel chromatography to obtain diester intermediate 1.07 inabout 52% yield. ¹H NMR (500 MHz, CDCl₃) δ: 5.34 (dd, J=8.1, 5.5 Hz,1H), 4.19 (q, J=7.1 Hz, 4H), 3.73-3.61 (m, 6H), 3.14 (dt, J=15.5, 5.5Hz, 1H), 3.02 (t, J=5.4 Hz, 2H), 2.28-0.64 (m, 49H, Cholesterolbackbone).

Step 7:

To a 50 mL single neck round bottom flask, diester compound 1.07 (about218 mg, 0.363 mmol) was taken in THF/water (about 4 mL, at a ratio ofabout 3:1) and cooled to about 0° C. To this cooled solution, LiOH(about 34 mg, 1.45 mmol) was added and stirred at room temperature for atime-period of another 6 h. After completion, the reaction mixture wasconcentrated under reduced pressure to remove THF and the aqueous layerwas washed with ethyl acetate. The aqueous layer was lyophilized to getsolid di-lithium salt of 1.08 with a quantitative yield.

Step 8: Synthesis of DACH-Pt(H₂O)₂

To a 50 mL single neck round bottom flask dichloro(1,2-diamino-cyclohexane) platinum 1.09 (about 200 mg, 0.526 mmol) wastaken in about 20.0 mL of H₂O. To this suspension, silver nitrate (about178.7 mg, 1.052 mmol) was added and the reaction mixture was stirred atroom temperature for about 24 h. The milky white solution wascentrifuged and the solution was filtered through 0.22 μM syringe filterto obtain aquated DACH-Pt 1.10 in quantitative yield (about 10 mg/mL).

Step 9:

To a 100 mL single neck round bottom flask intermediate 1.08 (about 202mg, 0.363 mmol) was taken in about 1.0 mL water. To this solution,DACHPt(H₂O)₂ (about 13.8 mL) obtained in the previous step was added andstirred for another 12 h. The solid residue was filtered and washed withwater (about 20 mL). The white solid residue was lyophilized anddissolved in excess methanol, filtered and concentrated under reducedpressure to afford cholesterol-oxaliplatin amphiphile Compound 25 inabout 85% yield. ¹H NMR of IO-125_01 (500 MHz, CDCl₃) δ: 5.36 (s, 1H),3.71 (s, 4H), 3.64 (m, 2H), 3.16 (m, 1H), 2.86-2.78 (m, 2H), 2.36-0.62(57H, cholesterol back bone). ¹³C NMR of IO-125_01 (125 MHz,CD₂Cl₂-CD₃OD) δ:183.13, 182.80, 171.64, 171.35, 140.13, 122.08, 79.93,56.73, 56.17, 50.19, 42.22, 39.75, 39.41, 38.87, 38.66, 37.00, 36.71,36.11, 35.75, 32.16, 31.84, 29.54, 28.24, 28.13, 28.08, 27.90, 24.34,24.19, 24.13, 23.72, 22.31, 22.06, 20.97, 18.97, 18.94, 18.32, 11.44; IRof IO-125_01 (KBr): 3416.28, 3162.69, 2933.20, 1654.62, 1599.66,1455.99, 1377.89, 1317.14, 1174.44, 1091.51, 1061.62; MALDI-TOF MS ofIO-125_01 C₃₉H₆₇N₃O₅Pt (m/z)=853.644 (M)⁺; ¹⁹⁵Pt of IO-125_01 (108 MHz,CD2Cl2-MeOD) −2316.5 and −2341.82; Analytical calculation found forC₃₉H₆₇N₃O₅Pt C, 52.63 (54.91), H, 7.93 (7.92), N, 4.21 (4.93).

Modified Synthesis of Compound 26

Step 1:

To an ice cooled solution of ethylene diamine (about 22.2 mL) in CH₂Cl₂(about 40 mL), a solution of compound 1.11 (about 5 g) in CH₂Cl₂ (about50 mL) was added dropwise over a period of about 45 min and the reactionmixture was stirred at the same temperature for about 1 h and wasfurther allowed to stir at room temperature for an additionaltime-period of about 20 h. After completion (checked by TLC), thereaction mixture was quenched with water and extracted with dichloromethane (about 4×50 mL), the combined organic layer was dried overanhydrous Na₂SO₄ and concentrated under vacuum. The residue was purifiedby column chromatography utilizing methanol-chloroform as mobile phaseto obtain intermediate 1.12. ¹H NMR (500 MHz, CDCl₃) δ: 5.30 (s, 1H),5.05 (s, 1H), 4.42 (s, 1H), 3.18 (s, 2H), 2.79 (s, 2H), 2.35-0.60 (m,45H, cholesterol backbone).

Step 2:

To a 50 mL single neck round bottom flask amine 1.12 (about 300 mg 0.634mmol) was taken in THF (about 5 mL) under nitrogen atmosphere. Thereaction mixture was cooled to about 0° C. under ice bath and NaH (about130 mg, 3.17 mmol) was added by pinch over a period of about 10 minutes.The resulting solution was stirred for about 20 minutes and ethyl bromoacetate was added. The reaction mixture was stirred for about 2 h atroom temperature and TLC was checked. After completion, the reactionmixture was cooled to about 0° C. and quenched with cold water (about 5mL), extracted with ethyl acetate (about 2×20 mL), dried over anhydrousNa₂SO₄ and thereafter concentrated. The residue was purified by columnchromatography utilizing methanol-chloroform as mobile phase to obtainintermediate 1.13.

¹H NMR (500 MHz, CDCl₃) δ: 5.85 (s, 1H), 5.39 (d, J=4.9 Hz, 1H), 4.50(m, 1H), 4.25-4.15 (m, 4H), 3.63 (m, 4H), 3.30 (bs, 2H), 2.98 (bs, 2H),2.44-0.71 (m, 49H, cholesterol backbone).

¹³C NMR (500 MHz, CDCl₃) δ: 171.32, 156.37, 140.00, 122.28, 74.14,60.79, 56.66, 56.09, 55.16, 53.36, 49.97, 42.28, 39.71, 39.49, 38.57,37.00, 36.54, 36.15, 35.77, 31.88, 31.85, 28.21, 28.14, 27.99, 24.26,23.79, 22.80, 22.55, 21.01, 19.32, 18.68, 14.19, 11.83;

Step 3:

To a 50 mL single neck round bottom flask, diester 1.13 (about 1.7 g,2.63 mmol) was taken in THF/water (about 3:1) (about 16 mL). Thereaction mixture was cooled to about 0° C. under ice bath and LiOH(about 130 mg, 5.27 mmol) was added to the reaction mixture. Theresulting solution was stirred for about 6 h at room temperature and TLCwas checked. After completion, the reaction mixture was concentratedunder reduced pressure to remove THF and diluted with water (about 5mL). The water layer was washed with ethyl acetate and CH₂Cl₂successively and lyophilized to obtain intermediate 1.14 in quantitativeyield.

Step 4:

To a 100 mL single neck round bottom flask, intermediate 1.14 was takenin about 1.0 mL water. To this solution, DACHPt(H₂O)₂ was added andstirred for a time-period of another 12 h. The solid residue obtainedwas filtered, washed with water and lyophilized. The residue wasdissolved in excess methanol, filtered and concentrated under reducedpressure to afford cholesterol-oxaliplatin amphiphile Compound 26. ¹HNMR of IO-126_01 (500 MHz, CDCl₃) δ: 5.40 (s, 1H), 4.89 (bs, 1H), 4.52(m, 1H), 3.61 (s, 4H), 3.40-3.28 (m, 2H), 2.73-2.66 (m, 2H), 2.44-0.60(57H, cholesterol back bone). ¹³C NMR of IO-126_01 (125 MHz,CD₂Cl₂-CD₃OD) δ: 186.87, 186.49, 175.25, 174.91, 161.48, 161.24, 143.69,126.49, 79.20, 60.62, 60.04, 53.92, 46.19, 43.62, 43.38, 42.4, 42.32,40.86, 40.46, 40.05, 39.68, 36.13, 35.80, 35.73, 33.97, 32.10, 31.96,31.87, 28.21, 28.14, 27.70, 26.58, 26.32, 23.09, 22.51, 15.66; IR ofIO-126 (KBr): 3403.74, 2931.27, 1665.23, 1632.45, 1444.42, 1382.71,1131.05; MALDI-TOF MS of IO-126_01 C₄₀H₆₈N₄O₆Pt (m/z)=896.5292 (M)⁺;¹⁹⁵Pt of IO-126_01 (108 MHz, MeOD) −2280.34 and −2305.06; Analyticalcalculation found for C₄₀H₆₈N₄O₆Pt C, 51.96 (53.62), H, 7.82 (7.65), N,5.39 (6.25).

Modified Synthesis of Compound 27

Step 1:

To a 50 mL single neck round bottom flask, BocHNCH₂COOH (about 370 mg,2.08 mmol) was taken in CH₂Cl₂ (about 10 mL) under nitrogen atmosphere.Solid EDCl (about 400 mg, 2.08 mmol) and HOBT (about 285 mg, 2.08 mmol)are added successively to the reaction mixture. DIPEA was added to makethe solution alkaline and the reaction mixture was stirred for another20 minutes. To this activated acid solution, amine 1.06 (about 450 mg,1.04 mmol) was added and the mixture was stirred at room temperature forabout 12 h and TLC was checked. After completion, the reaction mixturewas quenched with water, extracted with chloroform, dried over anhydrousNa₂SO₄ and thereafter concentrated. The residue was purified by silicagel chromatography utilizing methanol-chloroform as mobile phase toobtain intermediate 1.15.

Step 2:

To a 50 mL single neck round bottom flask, Boc protected amine 1.15(about 600 mg, 0.99 mmol) was taken in CH₂Cl₂ and the flask was cooledto about 0° C. To this solution, TFA was added and the mixture wasstirred for about 3 hours at the same temperature. After completion, thereaction mixture was concentrated under rotary evaporator and the crudeproduct 1.16 was utilized for the next reaction without furtherpurification.

Step 3:

To a 50 mL single neck round bottom flask, crude amine 1.16 (about 400mg, 0.821 mmol) was taken in THF (about 10 mL) under nitrogenatmosphere. The solution was cooled to about 0° C. under ice bath andsolid NaH (about 160 mg, 4.10 mmol) was added pinch wise over a periodof about 10 minutes. The resulting solution was stirred for anadditional 20 minutes and ethyl bromo acetate was added. Aftercompletion, the reaction mixture was cooled to about 0° C. and quenchedwith water, extracted with ethyl acetate, dried over anhydrous Na₂SO₄and thereafter concentrated under vacuum. The residue was purified bysilica gel chromatography utilizing methanol-chloroform as mobile phaseto obtain intermediate 1.17. ¹H NMR (500 MHz, CDCl₃) δ: 5.33-5.29 (m,1H), 4.19-4.12 (q, J=7.25 Hz, 4H), 3.56-3.49 (m, 6H), 3.43 (dd, J=11.6,5.9 Hz, 2H), 3.39 (s, 2H), 3.14 (m, 1H), 2.44-0.71 (m, 50H, cholesterolbackbone). ¹³C NMR (500 MHz, CDCl₃) δ: 170.90, 170.66, 140.77, 121.54,79.15, 66.33, 60.85, 58.82, 56.70, 56.09, 55.57, 50.12, 42.25, 39.71,39.45, 39.35, 38.95, 37.14, 36.79, 36.12, 35.71, 31.87, 31.83, 28.26,28.16, 27.93, 24.22, 23.75, 22.74, 22.49, 21.00, 19.29, 18.65, 14.15,11.79.

Step 4:

To a 50 mL single neck round bottom flask, diester 1.17 (about 200 mg,0.303 mmol) was taken in THF/water (about 3:1) (about 4 mL) at about 0°C. Solid LiOH (about 15 mg, 0.606 mmol) was added to the reactionmixture and stirred for about 6 hours at room temperature. Aftercompletion, the reaction mixture was concentrated and diluted with water(about 4 mL). The aqueous layer was washed with ethyl acetate anddichloro methane successively and was lyophilized to obtain solid powderof acid salt 1.18 in quantitative yield.

Step 5:

To a 100 mL single neck round bottom flask, intermediate 1.18 was takenin 1 mL water. To this solution, DACHPt(H₂O)₂ was added and stirred foranother 12 hours. The solid residue was filtered and washed with waterand thereafter lyophilized. The residue was dissolved in excessmethanol, filtered and concentrated under reduced pressure to affordcholesterol-oxaliplatin amphiphile Compound 27. ¹H NMR of IO-127_01 (500MHz, CDCl₃-CD₃OD) δ: 5.28 (s, 1H), 4.11 (d, J=14.1 Hz, 1H), 3.65-3.25(m, 8H), 3.17-3.05 (m, 1H), 2.44-0.60 (57H, cholesterol backbone). ¹³CNMR of IO-127_01 (125 MHz, DMSO d₆) δ:180.1, 168.19, 166.47, 140.36,121.06, 78.25, 65.47, 63.65, 61.23, 61.14, 60.31, 56.09, 55.48, 49.50,41.76, 38.54, 36.57, 36.19, 35.56, 35.09, 31.32, 31.27, 28.88, 27.94,27.68, 27.30, 23.77, 23.08, 22.58, 22.31, 20.51, 18.98, 18.46, 11.59; IRof IO-127_01 (KBr): 3447.72, 3245.64, 2935.69, 1617.40, 1392.53,1096.25; MALDI-TOF MS of IO-127_01 C₄₁H₇₀N₄O₆Pt (m/z)=910.6240 (M)⁺;¹⁹⁵Pt of IO-127_01 (108 MHz, MeOD) −2260.12 and −2271.67; Analyticalcalculation found for C₄₁H₇₀N₄O₆Pt C, 52.85 (54.11), H, 7.77 (7.75), N,5.26 (6.16).

Example 4: Synthesis of Compounds of Formula III Synthesis of Compound31

Step 1:

To a 50 mL single neck round bottom flask, acetonide protected keto-acid(about 121.5 mg, 0.854 mmol) is taken in about 2 mL anhydrous THF andthe mixture is cooled to about −78° C. To this solution, LiHMDS (about0.85 mL, 5 equiv, 1 mmol solution in toluene) is added and the mixtureis stirred for about 15 minutes at the same temperature. To thissolution, tosyl compound 1.04 (about 100 mg, 0.171 mmol) in THF (about 2mL) is added and again the mixture is allowed to stir for about 2 hoursat about −78° C. After completion, the reaction mixture is cooled toabout 0° C. and quenched with water and extracted with ethyl acetate(about 2×15 mL). The organic layer is dried over anhydrous Na₂SO₄,concentrated and purified by silica gel chromatography to obtain diesterintermediate 1.27.

Step 2:

Acetonide protected compound 1.27 in about 1 mL THF is taken in a 50 mLround bottom flask and HCl (about 1 M) at about 0° C. is added. Thereaction mixture is stirred for about 3 hours at room temperature andTLC is carried out. After completion, the reaction mixture is dilutedwith water (about 5 mL) and extracted with ethyl acetate (about 20 mL).The organic layer is dried over anhydrous Na₂SO₄, concentrated andpurified by silica gel chromatography to obtain intermediate 1.28.

Step 3:

To a 50 mL single neck round bottomed flask, hydroxyl acid A in DMF(about 1 mL) is taken and the mixture is stirred at room temperature forabout 30 minutes. Aquated DACH is added to the reaction mixture at roomtemperature and the mixture is stirred for about 24 hours and thereafterlyophilized. The solid residue is washed with water (about 5 mL) andlyophilized to achieve final platinum adduct product Compound 31.

Synthesis of Compound 32

Step 1:

To a 50 mL single neck round bottom flask, alcohol intermediate 1.03 istaken in about 5 mL anhydrous CH₂Cl₂ and cooled to about 0° C. To thissolution, (pyridinium chlorochromate) PCC is added and the reactionmixture is stirred at room temperature for about 3 hours and thereafter,the reaction is checked with TLC. After completion, the reaction mixtureis concentrated and purified by silica gel chromatography to obtainaldehyde intermediate.

Step 2:

To 50 mL single neck round bottom flask, TPP (Sodium triphosphate) saltin THF is taken and the mixture is cooled to about 0° C. To thissolution, nBuLi is added and the reaction mixture is stirred for about 1hour. To the solution obtained, the aldehyde prepared in the previousstep in THF (about 5 mL) is slowly added. The resulting solution isstirred for another 3 hours at about 0° C. and the progress of thereaction checked with TLC. After completion of the reaction, thereaction mixture is quenched with water and extracted with ethylacetate. The combined organic layer obtained is concentrated and purifyby silica gel chromatography.

Step 3:

To a 50 mL single neck round bottom flask, liquid ammonia in THF istaken at about −78° C. To this solution, metallic sodium is added slowlyover a period of about 20 minutes. To the obtained blue solution, benzylprotected compound in THF is added over a period of about 10 minutes.The resulting solution is stirred for about 3 hours at the sametemperature and the progress of the reaction is checked by TLC. Aftercompletion of the reaction, the reaction mixture is left for about 12hours at room temperature and thereafter, quenched with ammoniumchloride solution, extracted with ethyl acetate and finally concentratedunder reduced pressure. The residue is purified by silica gelchromatography.

Step 4:

To 50 mL single neck round bottom flask, hydroxyl compound in CH₂Cl₂ istaken at about 0° C. To this solution, solid Dess-Martin Periodinane(DMP) is added and the mixture is stirred for about 3 hours and thereaction is monitored by TLC. After completion of the reaction, thereaction mixture is quenched with water and extracted with CH₂Cl₂. Theorganic layer is thereafter concentrated and purified by silica gelchromatography.

Step 5:

To 50 mL single neck round bottom flask, lithium diisopropylamide (LDA)is added at about −78° C. To this, tertiary butyl acetate in THF isadded and the mixture stirred for about 0.5 hours. To this solution, thealdehyde prepared in the previous step in THF (about 5 mL) is slowlyadded. The resulting solution is stirred for another 2 hours at about−78° C. and the reaction is monitored by TLC. After completion, thereaction mixture is quenched with water and extracted with ethylacetate. The combined organic layer is concentrated and purified bysilica gel chromatography.

Step 6:

To 50 mL single neck round bottom flask, hydroxyl compound in CH₂Cl₂ istaken at about 0° C. To this solution, solid Dess-Martin Periodinane(DMP) is added and the mixture is stirred for about 3 hours and theprogress of the reaction is checked by TLC. After completion, thereaction mixture is quenched with water and extracted with CH₂Cl₂. Theorganic layer obtained is concentrated and purified by silica gelchromatography.

Step 7:

To a 50 mL single neck round bottom flask, tertiary butyl ester in THFis taken and the solution is cooled to about 0° C. To the cooledsolution, about 1 (M) HCl is added and the reaction mixture is stirredfor about 2 hours at the same temperature. After completion of thereaction, the compound is extracted with ethyl acetate and concentratedunder reduced pressure. The residue is purified by silica gelchromatography.

Step 8:

To a 50 mL single neck round bottomed flask, hydroxyl acid in DMF (about1 mL) is taken and the mixture is stirred at room temperature for about30 minutes. Aquated DACH is added to the reaction mixture at roomtemperature and stirred for about 24 hours and thereafter lyophilized.The solid residue is washed with water (about 5 mL) and lyophilized toobtain the final platinum adduct product Compound 32.

Synthesis of Compound 33

Step 1:

To a 50 mL single neck round bottom flask, alcohol intermediate 1.03 inabout 5 mL anhydrous CH₂Cl₂ is taken and cooled to about 0° C. To thissolution, PCC is added and the reaction mixture is stirred at roomtemperature for about 3 hours and the reaction progress is checked byTLC. After completion, the reaction mixture is concentrated andthereafter, purification is carried out by silica gel chromatography toobtain aldehyde intermediate.

Step 2:

In 50 mL single neck round bottom flask, LDA is generated at about −78°C. in THF. To this solution, 1, 3-dioxinones is added in THF and thereaction mixture is stirred for about 0.5 hours. To this solution,previously prepared aldehyde in THF (about 5 mL) is slowly added, andthe resulting solution is stirred for another 2 hours at about −78° C.and the reaction is checked by TLC. After completion, the reactionmixture is quenched with water and extracted with ethyl acetate. Thecombined organic layer is concentrated and thereafter purification iscarried out by silica gel chromatography.

Step 3:

To a 50 mL single neck round bottom flask, acetonide protected compoundin THF is taken at about 0° C. To this solution, 1 (M) HCl is added andthe reaction mixture is stirred for about 2 hours at same temperature.After completion of the reaction, the compound is extracted with ethylacetate and concentrated under reduced pressure. The residue is purifiedby silica gel chromatography.

Step 4:

To a 50 mL single neck round bottomed flask, hydroxyl acid in DMF (about1 mL) is taken and the solution is stirred at room temperature for about30 minutes. Aquated DACH-Pt(H₂O) is added to the reaction mixture atroom temperature and the reaction mixture is stirred for another 24hours and thereafter lyophilized. The solid residue is washed with water(about 5 mL) and thereafter lyophilized to obtain final platinum adductproduct Compound 33.

Synthesis of Compound 34 [Wherein, R=Cholesterol or Other Lipid]

Step 1:

To an ice cooled solution of cholesterol 1.01 in CH₂Cl₂, pyridine isadded and stirred for about 15 minutes. To this solution, p-toluenesulphonyl chloride is added and stirred for another 6 h at about 0° C.After completion, the reaction mixture is diluted with CHCl₃ and washedwith about 1N HCl and brine successively. The organic layer is driedover anhydrous Na₂SO₄ and concentrated under vacuum to affordintermediate 1.02 and the intermediate is employed for the next reactionwithout further purification.

Step 2:

To the solution of crude tosyl cholesterol 1.02 in dioxane, 1,3-propanediol is added and the reaction mixture is refluxed for about 4hours. After completion, the reaction mixture is extracted with ethylacetate and washed with water and brine successively. The organic layeris removed over anhydrous Na₂SO₄ and concentrated under vacuum andfinally the residue is purified on silica gel column to afford alcoholintermediate.

Step 3:

To an ice cooled solution of alcohol in CH₂Cl₂, pyridine is added andstirred for about 15 minutes. To this solution, p-toluene sulphonylchloride is added and the reaction mixture is stirred for another 6 h atabout 0° C. After completion, the reaction mixture is diluted with CHCl₃and washed with about 1N HCl and brine successively. The organic layeris dried over anhydrous Na₂SO₄ and thereafter concentrated under vacuum.The residue is purified on silica gel column to afford tosylintermediate.

Step 4:

In a 50 mL single neck round bottomed flask, methyl-3-mercaptopropionate in DMF (about 10 mL) is taken at about 0° C. under nitrogenatmosphere. Potassium carbonate is added to the reaction mixturefollowed by the addition of tosyl compound. The mixture is stirred atroom temperature for another 24 hours. After completion, the reactionmixture is quenched with water and thereafter extracted with ethylacetate. The organic layer is dried over anhydrous Na₂SO₄ andconcentrated under reduced pressure. The residue is finally purified bysilica gel chromatography to yield sulfide intermediate.

Step 5:

In a 50 mL single neck round bottom flask, ester compound in THF/wateris taken and the mixture is cooled to about 0° C. To this solution, LiOHis added and stirred at room temperature for another 3 hours. Aftercompletion, the reaction mixture is concentrated under reduced pressureto remove THF and further extracted with ethyl acetate. The organiclayer is dried over anhydrous Na₂SO₄, concentrated and finally purifiedby silica gel chromatography to obtain acid intermediate.

Step 6:

In a 50 mL single neck round bottom flask, the acid intermediateobtained in the previous step in CH₂Cl₂ is taken and the mixture iscooled to about 0° C. To this solution, m-chloroperoxybenzoic acid(mCPBA) (about 0.9 equivalents) is added and the reaction mixture isstir at the same temperature (i.e. 0° C.) for about 1 hour and theprogress of the reaction is checked by TLC. After completion of thereaction, the reaction mixture is quenched with water and furtherextracted with CH₂Cl₂. The organic layer is dried over anhydrous Na₂SO₄,concentrated and purified by silica gel chromatography to obtainpartially oxidized intermediate.

Step 7:

In a 50 mL single neck round bottom flask, acid intermediate in DMF istaken and the mixture is stirred at room temperature for about 15minutes. To this solution, DACHPt(H₂O)₂ is added and the reactionmixture is stirred for another 24 hours. The solution is lyophilized toafford amphiphile Compound 34 in good yield.

Synthesis of Compound 35 [Wherein, R=Cholesterol or Other Lipid]

Steps 1-5:

The sulphide intermediate obtained during the synthesis of compound 34(Steps 4 and 5) is taken as the starting reactant.

Step 6:

In a 50 mL single neck round bottom flask, the aforesaid sulphideintermediate in CH₂Cl₂ is taken and the mixture is cooled to about 0° C.To this solution, mCPBA (about 1.8 equivalent) is added and the reactionmixture is stirred at same temperature for about 1 hour and the reactionprogress is checked by TLC. After completion, the reaction mixture isquenched with water and thereafter extracted with CH₂Cl₂. The organiclayer is dried over anhydrous Na₂SO₄, concentrated and purified bysilica gel chromatography to obtain fully oxidized intermediate.

Step 7:

In a 50 mL single neck round bottom flask, acid intermediate in DMF istaken and stirred at room temperature for about 15 minutes. To thissolution, DACHPt(H₂O)₂ is added and the reaction mixture is stirred foranother 24 hours. The solution is lyophilized to afford amphiphileCompound 35 in good yield.

Synthesis of Compound 36 [wherein, R=Cholesterol or other lipid]

Steps 1-3:

The tosyl intermediate obtained during the synthesis of compound 34(Step 3) is taken as the starting reactant.

Step 4:

In a 50 mL round bottomed flask, the tosyl intermediate in DMF (about 20ml) is added under nitrogen atmosphere and stirred for about 30 minutesto get a clear solution (warming is carried out if necessary). To thissolution, sodium azide is added and the mixture is stirred for about 18hours at room temperature and TLC is employed to monitor the progress ofthe reaction. After completion of the reaction, the reaction mixture isdiluted with water, the compound is extracted with ethyl acetate,concentrated under vacuum and purified by flash chromatography to obtainazide intermediate.

Step 5:

To a solution of azide in dry DMF, triphenyl phosphene (TPP) is addedunder nitrogen atmosphere. The reaction mixture is stirred for about 6hours at room temperature and water is added to it. The reaction mixtureis again stirred at the same temperature for an additional 6 hours andTLC is employed to monitor the progress of the reaction. Aftercompletion of the reaction, organic solvent is removed under vacuum andthe residue is purified by silica gel chromatography usingmethanol/chloroform as eluent to obtain amine intermediate.

Step 6:

To an ice cool solution of amine in THF, NaH is added over a period ofabout 10 minutes under nitrogen atmosphere. The resulting solution isstirred for about 20 minutes and thereafter ethyl bromo acetate isadded. The reaction mixture is stir for another 6 hours at roomtemperature and TLC is employed to monitor the progress of the reaction.After completion of the reaction, the reaction mixture is cooled toabout 0° C. and quenched with water followed by extraction with ethylacetate. The organic layer is dried over anhydrous Na₂SO₄, concentratedand purified by silica gel chromatography to obtain ester intermediate.

Step 7:

To a 50 mL single neck round bottom flask, ester compound in THF/wateris taken and cooled to about 0° C. To this solution, LiOH is added andthe reaction mixture is stirred at room temperature for a time-period ofabout 3 hours. After completion of the reaction, the reaction mixture isconcentrated under reduced pressure to remove THF and extracted withethyl acetate. The organic layer is dried over anhydrous Na₂SO₄,concentrated and finally purified by silica gel chromatography to obtainacid intermediate.

Step 8:

In a 50 mL single neck round bottom flask, acid in CH₂Cl₂ is taken andcooled to about 0° C. To this solution, mCPBA (about 0.8 equivalents) isadded and the mixture is stirred at same temperature for about 1 hourand the reaction progress is monitored by TLC. After completion of thereaction, the reaction mixture is quenched with water and extracted withCH₂Cl₂. The organic layer is dried over anhydrous Na₂SO₄, concentratedand purified by silica gel chromatography to obtain N-oxideintermediate.

Step 9:

In a 50 mL single neck round bottom flask, N-oxide intermediate is takenin DMF and the mixture is stirred at room temperature for about 15minutes. To this solution, DACHPt(H₂O)₂ is added and the reactionmixture is stirred for a time-period of about 24 hours. The solution islyophilized to afford Compound 36 in good yield.

Example 5: Synthesis of Compound 30

Step 1:

In a 50 mL single neck round bottom flask, cholesterol 1.01 (about 1.0g, 2.59 mmol) in 5 mL anhydrous THF is taken under nitrogen atmosphereand cooled to about 0° C. To this solution, NaH (about 414 mg, 10.344mmol) is added and the mixture is stirred for about 30 minutes at thesame temperature (i.e. 0° C.). To this solution, ethyl bromo acetate(about 0.45 mL, 3.885 mmol) in THF (about 2 mL) is added and again thereaction mixture is allowed to stir for about 2 hours at roomtemperature. After completion, the reaction mixture is cooled to about0° C. and quenched with water followed by extraction with ethyl acetate(about 2×15 mL). The organic layer obtained is dried over anhydrousNa₂SO₄, concentrated and purified by silica gel chromatography to obtainester intermediate 1.29.

Step 2:

In a 50 mL single neck round bottom flask, ester 1.29 (about 220 mg,0.465 mmol) in THF/water (about 3:1) (about 4 mL) is taken at about 0°C. Solid LiOH (about 33 mg, 1.39 mmol) is added to the reaction mixtureand stirred for about 6 hours at room temperature. After completion ofthe reaction, the reaction mixture is acidified with saturated NaHSO₄ upto pH 3 followed by extraction with CHCl₃ (about 3×10 mL). The organiclayer is dried over anhydrous Na₂SO₄, concentrated under rotaryevaporator and purified by silica gel chromatography to obtain pure acidintermediate 1.30.

Step 3:

In a 50 mL single neck round bottomed flask, DACH(Cl)₂Pt (about 50 mg,0.131 mmol) in DMF (about 5 mL) is taken and stirred for about 10minutes. AgNO₃ (about 22 mg, 0.131 mmol) is added to the reactionmixture at room temperature and stirred for about 24 hours. Aftercompletion, the solid AgCl precipitate is removed by centrifugationfollowed by filtration through 0.2 micron syringe filter to obtain monochloro compound 1.31.

Step 4:

In a 50 mL single neck round bottomed flask, acid 1.30 in DMF (about 1mL) is taken and stirred at room temperature for about 30 minutes. Monochloro DACH platinum 1.31 is added to the reaction mixture at roomtemperature and stirred for about 24 hours. The solid residue is washedwith water (about 5 mL) and lyophilized to obtain the final platinumadduct product Compound 30.

Example 6: Synthesis of Exemplary Compounds Synthesis of Compound 63

Experimental Procedure

Compound A (1.0 mmol) is taken in 10 mL THF. To this, sulfoacetic acid(3.0 mmol) is added and the resulting solution is stirred for 24 hr atRT. The TLC is checked and after completion water is added to thereaction mixture and the unreacted A is extracted using ethyl acetate.Water layer used for next step.

Experimental Procedure

To a 50 mL single neck RBF aquated DACH platinum (0.1 mmol, 3 mL, 10mg/mL solution) is taken. Compound B (0.09 mmol), taken in 10 mL water,is added dropwise and the resulting solution is stirred at roomtemperature for 24 hrs. White precipitate appeared. Precipitate iswashed with water and dried over vacuum to obtain compound C.

Synthesis of Compound 64

Experimental Procedure

To a 25 mL single neck RBF compound B (1.0 mmol) (synthesized accordingto the procedure mentioned in compound 64a) is taken in 10 mL THF. Tothis, selenium (1.0 mmol) is added and the resulting solution is stirredfor 24 hr at RT. The TLC is checked and after completion water is addedto the reaction mixture and the unreacted A is extracted using ethylacetate. Water layer is used for next step.

Experimental Procedure

To a 50 mL single neck RBF aquated DACH platinum (0.1 mmol, 3 mL, 10mg/mL solution) is taken. Compound B (0.09 mmol), taken in 10 mL THF, isadded dropwise and the resulting solution was stirred at roomtemperature for 24 hrs. TLC is checked. THF evaporated to get a lightyellow precipitate. Precipitate is washed with water and dried overvacuum to obtain compound C.

Synthesis of Compound 65

Experimental Procedure

To a 25 mL single neck RBF compound B (1.0 mmol) (synthesized accordingto the procedure mentioned in compound 64a) is taken in 10 mL CCl₄. Tothis, chlorosulfonic acid (1.0 mmol) is added dropwise at 0° C. and theresulting solution is stirred for 24 hr at RT. TLC is checked and aftercompletion CCl₄ is evaporated in vac. 50 mL water is added and the crudeproduct is extracted in chloroform to obtain B as white powder.

Experimental Procedure

In a RBF B (0.13 mmol) in 5 mL (1:3 water: THF) is treated with 0.13mmol of sodium hydroxide at) 0° C. and the resulting solution is stirredfor 15 min. THF evaporated and water layer is added dropwise to thesolution of aquated platinum diaminocyclohexane (0.13 mmol in 15 mlwater). White precipitate formed during the reaction. Reaction mixturecentrifuged and precipitate was given water wash to get C as whitepowder.

Synthesis of Compound 37

Experimental Procedure

To a 25 mL single neck RBF compound B (1.0 mmol) (synthesized accordingto the procedure mentioned in compound 69) is stirred with phosphoricacid (H₃PO₃) (1.0 m mol), pyridine (5 mmol) and triethylamine (Et₃N) (2mmol) until clear solution is obtained. Acetic anhydride (2 mmol) isadded and the reaction mixture is stirred for 4 hrs at 80° C. After allB is consumed, as indicated by TLC, 5 mL water is added to the reactionmixture. Compound C is extracted by chloroform wash (25 mL×3). Solventconcentrated in vacuum to obtain C.

All successive steps to reach F are carried out according to theprocedure described for the preparation of IO-180_01

Synthesis of Compound 55

Experimental Procedure

A (compound A is synthesized according to the procedure mentioned in thesynthesis of compound 69). To a 25 mL single neck RBFdiphenylphosphinomethane (DPPM) (5.0 mmol) is taken in 30 mL THF. Tothis n-butyl-lithium (5.2 mmol) is added and the resulting solution isstirred for 15 min at 0° C. To the above solution cholesteryl bromide(A) (4.0 mmol) is added and the reaction is stirred for 16 hrs. The TLCwas checked and after completion water is added to the reaction mixtureand the compound is extracted using ethyl acetate. The combined organiclayer was concentrated under vacuum. Purified by column chromatography.

Experimental Procedure

To a 25 mL single neck RBF compound B (1.0 mmol) is taken in 30 mL THF.To this, hydrogen peroxide (2.2 mmol, 35% solution) is added and theresulting solution is stirred for 24 hr at RT. The TLC was checked andafter completion water is added to the reaction mixture and the compoundis extracted using ethyl acetate. The combined organic layer wasconcentrated under vacuum. Purified by column chromatography.

Experimental Procedure

To a 50 mL single neck RBF aquated DACH platinum (0.1 mmol, 3 mL, 10mg/mL solution) is taken. Compound C (0.09 mmol), taken in 10 mL THF, isadded dropwise and the resulting solution was stirred at roomtemperature for 24 hrs. TLC is checked. THF evaporated to get a lightyellow precipitate. Precipitate is washed with water and dried overvacuum to obtain compound E.

Synthesis of Compound 52

Experimental Procedure

To a 50 mL single neck RBF A (synthesis of A described in the ligandpreparation of compound 25) (1 mmol) was taken in 10 mL dry THF. Ph₂PCl(2 mmol) and triethylamine (2 mmol) are added. The reaction mixture isstirred under nitrogen for 12 hrs at RT. Solvent evaporated and columnchromatography is performed to obtain F.

Syntheses of F to Hare similar as described in the synthesis of compound55.

Experimental Procedure

To a 50 mL single neck RBF A (synthesis described in the ligandpreparation of compound 25) (1 mmol) was taken in 10 mL dry THF. Ph₂PCl(2 mmol) and triethylamine (2 mmol) are added. The reaction mixture isstirred under nitrogen for 12 hrs at RT. Solvent evaporated and columnchromatography is performed to obtain F.

Compound (G, 43) synthesis is similar as the synthesis of compound 44.

Synthesis of Compound 46, 47 and 48

Experimental Procedure

Compound (C, 46) synthesis is similar as the synthesis of compound 49.

Experimental Procedure

Compound (C, 47) synthesis is similar as the synthesis of compound 50.

Experimental Procedure: Compound (C, 48) synthesis is similar as thesynthesis of compound 50.

Synthesis of Compound 44 and 49

Experimental Procedure

A (compound A is synthesized according to the procedure mentioned in thesynthesis of compound 69). To a 25 mL single neck RBFdiphenylphosphinomethane (DPPM) (5.0 mmol) is taken in 30 mL THF. Tothis n-butyl-lithium (5.2 mmol) is added and the resulting solution isstirred for 15 min at 0° C. To the above solution cholesteryl bromide(A) (4.0 mmol) is added and the reaction is stirred for 16 hrs. The TLCwas checked and after completion water is added to the reaction mixtureand the compound is extracted using ethyl acetate. The combined organiclayer was concentrated under vacuum. Purified by column chromatography.

Synthesis of Compound 44:

To a 25 mL single neck RBF compound B (1.0 mmol) is taken in 30 mL THF.To this, aquated Pt(DACH) (1 mmol in 10 ml water) is added and theresulting solution is stirred for 24 hr at RT. THF is concentrated toobtain compound 44 as precipitate.

Experimental Procedure

To a 25 mL single neck RBF compound B (1.0 mmol) is taken in 30 mL THF.To this, hydrogen peroxide (1.2 mmol, 35% solution) is added and theresulting solution is stirred for 24 hr at RT. The TLC was checked andafter completion water is added to the reaction mixture and the compoundis extracted using ethyl acetate. The combined organic layer wasconcentrated under vacuum. Purified by column chromatography.

Experimental Procedure

To a 50 mL single neck RBF aquated DACH platinum (0.1 mmol, 3 mL, 10mg/mL solution) is taken. Compound C (0.09 mmol), taken in 10 mL THF, isadded dropwise and the resulting solution was stirred at roomtemperature for 24 hrs. TLC is checked. THF evaporated to get a lightyellow precipitate. Precipitate is washed with water and dried overvacuum to obtain compound E.

Synthesis of Compound 50

Experimental Procedure

To a 25 mL single neck RBF compound B (1.0 mmol) is taken in 30 mL THF.To this, sulfur (1.0 mmol) is added and the resulting solution isstirred for 24 hr at RT. The TLC is checked and after completion wateris added to the reaction mixture and the compound is extracted usingethyl acetate. The combined organic layer is concentrated under vacuum.Compound D purified by column chromatography.

Experimental Procedure

To a 50 mL single neck RBF aquated DACH platinum (0.1 mmol, 3 mL, 10mg/mL solution) is taken. Compound C (0.09 mmol), taken in 10 mL THF, isadded dropwise and the resulting solution was stirred at roomtemperature for 24 hrs. TLC is checked. THF evaporated to get a lightyellow precipitate. Precipitate is washed with water and dried overvacuum to obtain compound D.

Synthesis of Compound 51

Experimental Procedure

To a 25 mL single neck RBF compound B (1.0 mmol) is taken in 30 mL THF.To this, selenium (1.0 mmol) is added and the resulting solution isstirred for 24 hr at RT. The TLC is checked and after completion wateris added to the reaction mixture and the compound is extracted usingethyl acetate. The combined organic layer is concentrated under vacuum.Compound D purified by column chromatography.

Experimental Procedure

To a 50 mL single neck RBF aquated DACH platinum (0.1 mmol, 3 mL, 10mg/mL solution) is taken. Compound C (0.09 mmol), taken in 10 mL THF, isadded dropwise and the resulting solution was stirred at roomtemperature for 24 hrs. TLC is checked. THF evaporated to get a lightyellow precipitate. Precipitate is washed with water and dried overvacuum to obtain compound D.

Synthesis of Compound 54

Experimental Procedure

Same as 50, two equivalent of sulfur is used.

Experimental Procedure

Same as 51, two equivalent of selenium is used.

Synthesis of Compound 57

Experimental Procedure

Same as 50, two equivalent of sulfur is used.

Experimental Procedure

Same as 51, two equivalent of selenium is used.

Synthesis of Compound 58, 59, 60

Experimental Procedure

To a 50 mL single neck RBF, K₂PtCl₄ (0.1 mmol, 3 mL, 10 mg/mL solution)is taken. Compound A (0.1 mmol), taken in 10 mL THF, is added dropwiseand the resulting solution is stirred at room temperature for 24 hrs.TLC is checked. THF evaporated to get a light yellow precipitate.Precipitate is washed with water and dried over vacuum to obtaincompound D.

Synthesis of Compound 61 and 62

Experimental Procedure

To a 50 mL single neck RBF, compound B (1 mmol, in 10 ml DMF) is taken.Silver nitrate (2 mmol) is added and stirred for 24 hrs. Whiteprecipitate is separated by filtration. Compound C (1 mmol), taken in 10mL water, is added dropwise to the filtrate and the resulting solutionis stirred at room temperature for 24 hrs. TLC is checked. DMFevaporated to get a light yellow precipitate. Precipitate is washed withwater and dried over vacuum to obtain compound 61/62.

Synthesis of Compound 78

Compound A (0.5 mmol) is taken in 20 ml water and silver nitrate (1mmol) is added. Reaction mixture is stirred for 24 hrs, filtered andDMSO (0.5 mmol) is added. Reaction mixture is stirred at roomtemperature for 2 hrs to obtain a yellow precipitate as compound 78.

Synthetic Scheme to Obtain Compound 79

Synthetic Scheme to Obtain Compound 80

Synthetic Scheme for Compound 81

Synthetic Scheme for Compound 82

Synthetic Scheme for Compound 83

Synthetic Scheme for Compound 84

Synthetic Scheme for Compound 85

Synthesis of Compound 95

Compound A (1 mmol) is taken in a 50 ml rb with 10 ml THF. AgBF₄ (1mmol) is added and stirred at room temperature for 24 hrs. Precipitateis filtered and compound B (1 mmol in 10 ml water) is added to thefiltrate dropwise. After 24 hrs of RT stirring, THF evaporated and theprecipitate obtained is filtered and washed with water to get compound95.

Example 7: Synthesis of Additional Exemplary Compounds Synthesis ofIO-131

Experimental Procedure

To the solution of tosylated cholesterol A (7.4 mmol) in dioxane (50 mL)was added ethylene glycol (35 mL) and refluxed for 4 h. The TLC waschecked. After completion the reaction mixture was concentrated undervacuum to remove dioxane and then it was extracted with ethyl acetateand washed with water (3×50 mL) and brine (20 mL) successively. Theorganic layer was dried over anhydrous Na₂SO₄ and concentrated undervacuum and column was performed.

Experimental Procedure

To the solution of cholesteryl ethylene glycol B (4.5 mmol) in DCM 5 mlwas added triphenyl phosphine (TPP) (9 mmol) and carbon tetra bromide (9mmol). The reaction mixture was stirred for 6 h at rt and TLC waschecked. After completion the reaction mixture was diluted with CHCl₃(20 mL) and washed with water (3×50 mL) and brine (20 mL) successively.The organic layer was dried over anhydrous Na₂SO₄ and concentrated undervacuum and put for flash column to get the pure compound (C).

Experimental Procedure

To a 25 mL single neck RBF methyl-3-mercaptopropionate C (5.0 mmol) wastaken in 30 mL DMF. To this potassium carbonate (20.0 mmol) was addedand the resulting solution was stirred for 15 minutes at RT. To theabove solution cholesteryl bromide (B) (4.0 mmol) was added and thereaction was stirred for 16 hrs. The TLC was checked and aftercompletion water was added to the reaction mixture and the compound wasextracted using ethyl acetate. The combined organic layer wasconcentrated under vacuum. Purified by column chromatography.

Experimental Procedure

To a 50 mL single neck RBF compound D (1.87 mmol) was taken in 60 mLDCM. To this mCPBA (1.31 mmol) was added and the resulting solution wasstirred for 3 hrs at 0° C. The TLC was checked and after completionwater was added to the reaction mixture and the compound was extractedusing chloroform. The combined organic layer was concentrated undervacuum and put for column chromatography. ¹H NMR (CDCl₃): 0.66 (s), 0.84to 0.47 (m), 1.82-1.97 (m), 2.21 (m), 2.35 (m), 2.84 (m), 2.98 (m), 3.16(t), 3.21 (m), 3.89 (br, s), 5.33 (br, s). ¹³C NMR (CDCl₃): 11.81,18.68, 19.32, 21.03, 22.52, 22.78, 23.78, 24.25, 26.96, 27.97, 28.18,31.84, 35.74, 36.15, 36.79, 37.05, 38.78, 38.91, 39.47, 39.72, 42.27,47.18, 50.10, 52.14, 53.25, 53.29, 56.11, 56.71, 79.71, 121.90, 140.40,171.74. Mass-ESI: 571.4 (M+Na) IR (KBr) (ν, cm⁻¹): 418 (w), 668 (m), 750(m), 1020 (m), 1104 (w), 1134 (w), 1178 (w), 1259 (m), 1275 (m), 1455(m), 1732 (m), 2933 (s), 3612 (m), 3723 (m), 3852 (m)

Experimental Procedure

To a 50 mL single neck RBF ester E (0.17 mmol) was taken in 3 mL ofTHF/H₂O (3:1) and cooled to 0° C. under ice bath. To this ice cooledsolution KOH (0.19 mmol in 2 mL) was added and was stirred at RT for 12h, the TLC was checked. After completion, THF was evaporated andremaining was extracted using chloroform. Water layer was used for nextstep.

Experimental Procedure: To a 50 mL single neck RBF aquated DACH platinum(3 mL, 10 mg/mL solution) was taken. Salt A (0.09 mmol), taken in 10 mLwater was added dropwise and the resulting solution was stirred at roomtemperature for 24 hrs. White precipitate separated and washed with 30ml of water to get the pure compound (G). IR (KBr) (ν, cm⁻¹): 415 (w,br), 797 (w), 1024 (m), 1107 (m), 1259 (w), 1377 (m), 1466 (w, br), 1588(m, br), 2933 (s), 3176 (w, br), 3440 (w, br). ¹⁹⁵Pt NMR(CDCl₃): −2893.

Synthesis of IO-148_01

Synthesis of Tosyl Intermediate of IO-148_01

Experimental Procedure

To an ice cooled solution of cholesterol A (10 g, 25.883 mmol) in CH₂Cl₂(150 mL) pyridine (10.5 mL, 129.415 mmol) was added dropwise and stirredfor 15 minutes. To the above solution p-toluene sulphonyl chloride B(14.75 g, 77.649 mmol) was added and stirred for 2 h under darkconditions. The TLC was checked and after completion of the reaction theorganic phase was washed with 0.1 N HCl solution (5×50 mL) and water(2×50 mL); the organic layer was dried over anhydrous Na₂SO₄ andconcentrated under vacuum.

Synthesis of Glycol Intermediate of IO-148_01

Experimental Procedure

To the solution of tosylated cholesterol A (10 g, 18.48 mmol) in dioxane(50 mL) was added ethylene glycol (35 mL) and refluxed for 4 h. The TLCwas checked. After completion the reaction mixture was concentratedunder vacuum to remove dioxane and then it was extracted with ethylacetate and washed with water (3×50 mL) and brine (20 mL) successively.The organic layer was dried over anhydrous Na₂SO₄ and concentrated undervacuum and column was performed.

Synthesis of Glycol Tosyl Intermediate of IO-148_01

Experimental Procedure

To an ice cooled solution of cholesteryl ethylene glycol A (6 g, 13.93mmol) in DCM 30 ml under nitrogen atmosphere was added p-toluenesulphonyl chloride B (3.25 g, 16.71 mmol) and stirred for 15 minutes. Tothis solution pyridine (12 mL) was added and stirred for 6 h at 0° C.and TLC was checked. After completion the reaction mixture was dilutedwith CHCl₃ (20 mL) and washed with saturated CuSO₄ (3×50 mL) and brine(20 mL) successively. The organic layer was dried over anhydrous Na₂SO₄and concentrated under vacuum.

Synthesis of azide intermediate of IO-148_01

Experimental Procedure

To the compound A (7 g, 11.97 mmol) DMF 20 ml was added under nitrogenatmosphere was stirred for 30 minutes to get a clear solution warm ifnecessary. To this solution sodium azide B 1.5 g, 23.95 mmol) was addedat once and stirred for 18 h at rt and TLC was checked. After completionthe reaction mixture was quenched with water and extracted with ethylacetate. The organic layer was given water wash and the combined organiclayer was concentrated under vacuum.

Synthesis of amine intermediate of IO-148_01

Experimental Procedure

To the Azide A (5 g, 10.97 mmol) dry THF (20 ml) was added undernitrogen atmosphere and TPP (5.74 g, 21.94 mmol) was added. The reactionwas stirred for 6 hr. After that 2 mL of water was added to the reactionmixture and reaction was kept at same temperature overnight. The TLC waschecked and the after completion of reaction, the reaction mixture wasconcentrated under vacuum and directly loaded to column.

Synthesis of N-Monoalkyl Intermediate of IO-148_01

Experimental Procedure

To a 50 mL single neck R.B flask amine A (200 mg, 0.465 mmol) was takenin anhydrous DCM (40 mL) under nitrogen atmosphere at 0° C. To thiscooled solution DIEPA (0.06 mL, 0.372 mmol) was added dropwise andstirred at same temperature for 20 minutes. To the above mixtureethylbromoacetate (0.03 mL, 0.279 mmol, in 10 mL DCM) was added dropwiseover a period of 1 hrs. Reaction was monitored using TLC. Aftercompletion the reaction mixture was directly concentrated under vacuumand put for Column chromatography.

Synthesis of N-Oxide Intermediate of IO-148_01

Experimental Procedure

To a 50 mL single neck R.B flask ester A (100 mg, 0.193 mmol) was takenin anhydrous CH₂Cl₂ (10 mL) under nitrogen atmosphere at 0° C. To thiscooled solution mCPBA (16.72 mg, 0.135 mmol) was added dropwise as asolution in DCM (2 mL) and stirred at same temperature for 2 h. Reactionwas monitored using TLC. After completion the reaction mixture wasquenched with NaHCO₃, extracted with CHCl₃, dried over anhydrous sodiumsulphate, concentrated and directly put for column chromatography.

Synthesis of Final Ligand of IO-148_01

Experimental Procedure

To a 50 mL single neck R.B flask N-oxide intermediate A (100 mg, 0.188mmol) was taken in anhydrous THF/H₂O (4 mL) at 0° C. To this cooledsolution LiOH.H₂O (8 mg, 0.188 mmol) was added and stirred at sametemperature for 1 h. Reaction was monitored using TLC. After completionthe reaction mixture was concentrated under vacuum to remove THF. Theresidue was diluted with water 10 mL and washed with DCM and ethylacetate.

Synthesis of IO-148_01

Experimental Procedure

To a 50 mL single neck RBF aquated DACH Platinum (70 mg, 0.188 mmol, 10mg/mL solution) was taken. To the above solution ligand A (100 mg, 0.188mmol) in 10 mL water was added dropwise. The resulting solution wasstirred for 3 h at room temperature. After completion the reactionmixture was centrifuged to separate the precipitate. The precipitateswere washed with water twice (10 mL) and lyophilized to get IO-148_01.ESIMS m/z=827.4

Synthesis of IO-148_02

Step 1-5 are similar as IO-148_01:

Step 6: Synthesis of N-methyl intermediate of IO-148_02

Experimental Procedure

To a 50 mL single neck R.B flask ester A (1 g, 1.93 mmol) was taken inacetonitrile under nitrogen atmosphere at 0° C. To the above mixturemethyl iodide (273 mg, 1.93 mmol) was added. Reaction was monitoredusing TLC. After completion the reaction mixture was directlyconcentrated under vacuum and put for Column chromatography.

Step 7: Synthesis of N-oxide intermediate of IO-148_02

Experimental Procedure

To a 50 mL single neck R.B flask ester A (100 mg, 0.193 mmol) was takenin anhydrous CH₂Cl₂ (10 mL) under nitrogen atmosphere at 0° C. To thiscooled solution mCPBA (16.72 mg, 0.135 mmol) was added dropwise as asolution in DCM (2 mL) and stirred at same temperature for 2 h. Reactionwas monitored using TLC. After completion the reaction mixture wasquenched with NaHCO₃, extracted with CHCl₃, dried over anhydrous sodiumsulphate, concentrated and directly put for column chromatography.

Step 8: Synthesis of Final Ligand of IO-148_02

Experimental Procedure

To a 50 mL single neck R.B flask N-oxide intermediate A (100 mg, 0.188mmol) was taken in anhydrous THF/H₂O (4 mL) at 0° C. To this cooledsolution LiOH.H₂O (8 mg, 0.188 mmol) was added and stirred at sametemperature for 1 h. Reaction was monitored using TLC. After completionthe reaction mixture was concentrated under vacuum to remove THF. Theresidue was diluted with water 10 mL and washed with DCM and ethylacetate.

Step 9: Synthesis of IO-148_02

Experimental Procedure

To a 50 mL single neck RBF aquated DACH Platinum (70 mg, 0.188 mmol, 10mg/mL solution) was taken. To the above solution ligand A (100 mg, 0.188mmol) in 10 mL water was added dropwise. The resulting solution wasstirred for 3 h at room temperature. After completion the reactionmixture was centrifuged to separate the precipitate. The precipitateswere washed with water twice (10 mL) and lyophilized to get IO-148_01.

Synthesis of IO-183_01

Experimental Procedure

To an ice cooled solution of cholesterol A (5 g, 12.93 mmol) in CH₂Cl₂(35 mL) was added pyridine (5.22 mL) and stirred for 15 minutes. To thissolution p-toluene sulphonyl chloride B (6.15 g, 32.31 mmol) was addedand stirred for 6 h at 0° C. and TLC was checked. After completion thereaction mixture was diluted with CHCl₃ (20 mL) and washed with 1N HCl(3×50 mL) and brine (20 mL) successively. The organic layer was driedover anhydrous Na₂SO₄ and concentrated under vacuum. Withoutpurification, the whole compound is used for the next reaction.

Experimental Procedure

To the solution of tosylated cholesterol A (10 g, 18.49 mmol) in dioxane(30 mL) was added diethylene glycol (10 mL) and refluxed for 4 h. TheTLC was checked. After completion the reaction mixture was extractedwith ethyl acetate and washed with water (3×50 mL) and brine (20 mL)successively. The organic layer was dried over anhydrous Na₂SO₄ andconcentrated under vacuum and column was performed. (Yield 38%)

Experimental Procedure

To a 100 mL single neck RBF NaH (594 mg) was taken in THF (10 mL) undernitrogen atmosphere. The reaction was cooled to 0° C. under ice bath andto it solution of C (2.35 g, 4.95 mmol) in THF (15 mL) was added slowly.The resulting solution was stirred for 1 h and ethyl bromo acetate wasadded slowly and stirred for 6 h at room temperature and TLC waschecked. After completion the reaction mixture was cooled to 0° C. andquenched with water, extracted with ethyl acetate. The organic layer waswashed with water and dried over Na₂SO₄ and concentrated and thecompound was purified by column chromatography. (Yield 46%)¹H NMR(CDCl₃): 0.66 (s), 0.81-2.41 (m), 3.2 (br, s), 3.6-3.8 (m), 4.36 (s),5.33 (s) ¹³C NMR (CDCl₃): 11.83, 18.68, 19.34, 21.03, 22.53, 22.39,23.79, 24.26, 27.58, 28.21, 29.67, 31.84, 31.91, 35.35, 36.15, 36.82,37.15, 38.89, 39.68, 39.75, 42.28, 50.12, 56.11, 56.73, 67.01, 68.76,70.11, 70.95, 71.41, 79.64, 121.67, 140.74, 172.69.

Experimental Procedure

To a 100 mL single neck RBF ester D (0.15 g, 0.28 mmol) was taken in 2mL of THF/H₂O (3:1) and cooled to 0° C. under ice bath. To this icecooled solution LiOH (12 mg, 0.28 mmol) was added and was stirred at rtfor 2 h, the TLC was checked. After completion the reaction mixture, THFwas removed by rotavapor. Chloroform was added to the reaction mixture.Compound was extracted with water. Then whole reaction mixture was usedfor the next reaction after rotavapor treatment.

Experimental Procedure

To a 50 mL single neck RBF DACH platinum (78 mg, 0.139 mmol) was takenin 5 mL HPLC Water. To the above solution silver nitrate (47 mg, 0.278mmol) was added. The resulting solution was stirred under protectionfrom light at rt. After 24 h, AgI precipitate was filtered. Filtrate wasused for the next step.

Experimental Procedure

To a 100 mL single neck RBF salt E (150 mg, 0.263 mmol) was taken in 40mL HPLC water and the resulting solution was stirred for 5 min at roomtemperature and to this solution DACH (OH₂)₂ platinum B was added and itwas stirred under protection from light at rt for 24 h. The precipitatewas filtered through filter paper and simultaneously washed with HPLCwater, HPLC Methanol and HPLC acetone and dried. (Yield 45%) ESIMSm/z=1395.7 [M+Na]⁺ for C₇₂H₁₂₄N₂O₁₀P ¹H NMR: (500 MHz, CDCl₃): 5.96(bs), 5.32 (s), 4.96 (bs), 3.92 (q), 3.61 (s), 3.15 (m), 2.57 (m), 2.37(m), 2.21 (t), 1.91 (m), 1.48 (m), 1.32 (m), 1.24 (m), 1.11 (m), 0.98(s), 0.90 (d), 0.85 (dd), 0.66 (s) ppm. ¹³C NMR (500 MHz, CDCl₃):177.22, 140.86, 121.56, 79.58, 70.70, 70.37, 70.31, 70.07, 67.26, 62.24,56.75, 56.17, 50.15, 42.30, 39.77, 39.50, 39.08, 37.23, 36.85, 36.18,35.79, 32.04, 31.94, 31.88, 28.36, 28.22, 27.98, 24.62, 24.28, 23.85,22.79, 22.54, 21.06, 19.38, 18.71, 11.85 ppm. IR: 418 (w), 668 (br, s),749 (m), 1110 (s), 1260 (s), 1640 (s), 2064 (w), 2933 (m), 3446 (br, s)

Synthesis of IO-183_02

Step 1:

Experimental Procedure

A (same procedure followed from ref. PNAS; 109, 2012; 11294) (200 mg,0.349 mmol) was dissolved in 2.5 mL of THF and 0.8 mL water. To it 16 mgof LiOH was added and stirred for 24 h at RT. White suspension appeared.THF was evaporated under vacuum and 40 mL of water is added to dissolvethe white residue. Water solution was washed with chloroform. Waterlayer used for next step.

Experimental Procedure

0.17 mmol cyclohexyldiamineplatinum-dichloride, 0.34 silver nitrate and7 mL water were added in a 25 mL RB and stirred for 48 h at room temp.Solution was centrifuged (4,000 rpm; 10 min) and white precipitate wasfiltered through syringe filter (25 mm/0.20 μm). Washed with 2 mL ofwater. Filtrate used for next reaction.

Step 3

Experimental Procedure

Compound B (0.26 mmol) in 20 ml water was added dropwise to C (0.13mmol) in water (10 ml). Reaction continued at room temperature for 20 h.White precipitate was separated by filtration. Residue washed with water(10 ml) to get D as white powder. ¹H NMR (CDCl₃+CD₃OD): 0.66 (s),0.84-2.5 (br, m), 3.32 (br, d), 4.45 (br, s), 5.34 (br, s) ¹³C(CDCl₃+CD₃OD) NMR: 11.68, 18.53, 19.15, 20.89, 22.36, 22.62, 23.7,24.12, 24.32, 27.84, 28.02, 29.53, 35.65, 36.03, 36.41, 38.36, 38.42,39.27, 39.36, 39.58, 40.27, 42.16, 49.56, 49.88, 56.02, 56.55, 62.44,74.46, 122.42, 139.63, 156.91, 174.20, 180.89 ESIMS: 1475.4 (M+Na) IR:418 (m), 584 (m), 799 (s), 1027 (m), 1260 (s), 1454 (m), 1535 (s), 1643(s), 1700 (br, s), 2928 (s), 3418 (br, s).

Synthesis of IO-147_02

Synthesis of IO-147_02:

Experiment Procedure

Compound B (synthesis described in the preparation of compoundIO-183_01) (1 mmol), phosphoric acid (1 mmol), pyridine (5 mmol) andtrimethylamine (2 mmol) are added in a round bottom flask (RBF) andstirred until clear solution appear. Acetic anhydride (2 mmol) is addeddropwise and the reaction mixture is stirred for 3 hrs at 80° C. Cooledto room temperature and water is added. Compound is extracted bydiethylether and concentrated under vacuum to obtain compound C. Rest ofthe reactions to obtain F is similar as described for the synthesis ofIO-180_01.

Synthesis of IO-173_01

Experimental Procedure

To an ice cooled solution of cholesterol A (5 g, 12.93 mmol) in CH₂Cl₂(35 mL) was added pyridine (5.22 mL) and stirred for 15 minutes. To thissolution p-toluene sulphonyl chloride B (6.15 g, 32.31 mmol) was addedand stirred for 6 h at 0° C. and TLC was checked. After completion thereaction mixture was diluted with CHCl₃ (20 mL) and washed with 1N HCl(3×50 mL) and brine (20 mL) successively. The organic layer was driedover anhydrous Na₂SO₄ and concentrated under vacuum. Withoutpurification, the whole compound is used for the next reaction.

Experimental Procedure

To the solution of tosylated cholesterol A (10 g, 0.018 mol) in dioxane(25 mL) was added ethylene glycol (15 mL) and refluxed for 4 h. The TLCwas checked. After completion the reaction mixture was extracted withethyl acetate and washed with water (3×50 mL) and brine (20 mL)successively. The organic layer was dried over anhydrous Na₂SO₄ andconcentrated under vacuum and it was purified by silica gelchromatography. (Yield=37%)

Experimental Procedure

A mixture of Compound B (1829 mg, 4.25 mmol) and Meldrum's acid A (612mg, 4.25 mmol) in anhydrous 1,4-dioxane (40 mL) was heated at 110° C.for 4 h. After cooling to room temperature, the reaction mixture waspartitioned with ethyl acetate and water. The organic extract was driedon Na₂SO₄ and concentrated using rotavapor and it was purified by silicagel chromatography. (Yield=26%)¹H NMR (500 MHz, CDCl₃): 5.33 (s), 4.26(s), 3.68 (s), 3.42 (s), 3.17 (s), 2.32 (d), 2.17 (s), 1.99 (m), 1.86(m), 1.48 (m), 1.33 (m), 1.24 (m), 1.11 (m), 0.98 (m), 0.90 (m), 0.85(m), 0.66 (s) ppm. ¹³C NMR (500 MHz, CDCl₃): 169.85, 167.25, 140.57,121.84, 79.70, 65.43, 65.27, 56.73, 56.13, 50.13, 42.29, 40.48, 39.74,39.49, 38.86, 37.13, 36.80, 36.16, 35.76, 31.91, 31.85, 29.67, 28.21,27.98, 24.26, 23.80, 22.79, 22.54, 21.04, 19.33, 18.89, 11.84 ppm.

Experimental Procedure

To a 50 mL single neck RBF DACH platinum A (100 mg, 0.263 mmol) wastaken in 10 mL HPLC Water. To the above solution silver nitrate (89 mg,0.526 mmol) was added. The resulting solution was stirred underprotection from light at rt. After 24 h, AgCl precipitate was filtered.Filtrate was used for the next step.

Experimental Procedure

To a 50 mL single neck RBF Acid B (136 mg, 0.263 mmol) was taken in 15mL Dry THF. To the above solution, DACH(OH₂)₂ platinum A (95 mg, 0.263mmol) was added dropwise under protection from light and stirred for 24h. Then the whole THF was evaporated. Precipitate was filtered and thewater part was lyophilized. ESIMS (M 824).

Synthesis of IO-173_03

Experimental Procedure

To an ice cooled solution of ethylene diamine B (22.2 mL) in 40 mL. DCMwas added solution of compound A (5 g) in DCM (50 mL) dropwise over aperiod of 45 min and stirred at the same temperature for 1 h and left atrt for additional 20 h. The TLC was checked and after completion of thereaction was quenched with water (4×100 mL) and the organic layer wasextracted with DCM (2×50 mL) and was dried over anhydrous Na₂SO₄ andconcentrated under vacuum and purified by silica gel columnchromatography. Yield 90%.

Experimental Procedure

A mixture of Compound A (2.74 g, 5.8 mmol) and Meldrum's acid A (661 mg,5.8 mmol) in anhydrous 1,4-dioxane (20 mL) was heated at 110° C. for 4h. After cooling to room temperature, the reaction mixture waspartitioned with ethyl acetate and Water. The organic extract was driedon Na₂SO₄ and concentrated using rotavapor. It was purified by silicagel column chromatography. Yield 50%.

Experimental Procedure

To a 50 mL single neck R.B sodium hydride (620 mg, 15.516 mmol) wastaken in THF (5 mL) under nitrogen atmosphere. The reaction mixture wascooled to 0° C. under ice bath and cholesterol A (2.82 g, 5.172 mmol) inTHF (10 mL) was added dropwise to the reaction mixture over a period of10 minutes and it was left for 30 min stirring. To this solution MethylIodide (2.42 g, 15.516 mmol) was added slowly and stirred for 6 h atroom temperature and TLC was checked. After completion the reactionmixture was cooled to 0° C. and quenched with water and extracted withethyl acetate, dried over anhydrous Na₂SO₄, concentrated and purified bysilica gel chromatography to obtain ester E in 40% yield.

Experimental Procedure

To a 50 mL single neck RBF ester A (0.154 g, 0.263 mmol) was taken in 2mL of THF/H₂O (3:1) and cooled to 0° C. under ice bath. To this icecooled solution LiOH B (11 mg, 0.263 mmol) was added and was stirred atrt for 3 h, the TLC was checked. After completion the reaction mixture,THF was removed by rotavapour. Chloroform was added to the reactionmixture. Compound was extracted with water. Then whole reaction mixturewas used for the next reaction after rotavapour treatment.

Experimental Procedure

To a 50 mL single neck RBF DACH platinum A (100 mg, 0.263 mmol) wastaken in 10 mL HPLC Water. To the above solution silver nitrate (88 mg,0.526 mmol) was added. The resulting solution was stirred underprotection from light at rt. After 24 h, AgCl precipitate was filtered.Filtrate was used for the next step.

Experimental Procedure

To a 100 mL single neck RBF Acid A (154 mg, 0.263 mmol) was taken in 20mL HPLC water and the resulting solution was stirred for 5 min at roomtemperature and to this solution DACH (OH₂)₂ platinum B was added and itwas stirred under protection from light at rt for 24 h. The precipitatewas filtered through filter paper and simultaneously washed with HPLCwater, HPLC Methanol and HPLC acetone and dried. (Yield 47%).

Synthesis of IO-176_01

Experimental Procedure

To a 50 mL single neck RBF DACH platinum A (100 mg, 0.263 mmol) wastaken in 20 mL HPLC Water. To the above solution silver nitrate (44 mg,0.263 mmol) was added. The resulting solution was stirred underprotection from light at rt. After 24 h, AgCl precipitate was filtered.Filtrate was used for the next step.

Experimental Procedure

To a 100 mL single neck RBF Acid A (same ligand as LB 55c) (154 mg,0.263 mmol) was taken in 20 mL HPLC water and the resulting solution wasstirred for 5 min at room temperature and to this solution DACH (OH₂)₂platinum B (0.263 mmol) was added and it was stirred under protectionfrom light at rt for 24 h. The precipitate was filtered through filterpaper and simultaneously washed with HPLC water, HPLC Methanol and HPLCacetone and dried. (Yield 40%)

Synthesis of IO-179_01

Step-1

Experimental Procedure

To a 50 mL single neck RBF aminomethylphosphonic acid A (0.77 mmol) ismixed with 2 mL dry pyridine. Cholesterol (0.77 mmol) and DMAP (0.77mmol) are added to the mixture and the resulting solution is stirred for16 h at RT. Resulting solution is acidified by dilute sulfuric acid andcompound C is extracted by chloroform washing.

Step-2

Experimental Procedure

To a 25 mL single neck RBF C (0.13 mmol) is taken in 1 mL THF. To thissolution KOH (0.26 mmol) in 1 ml water is added at 0° C. Immediate pptappeared. 2 ml water is added to dissolve the ppt and the resultingsolution is stirred for 2 h at RT. Reaction mixture is given chloroformwash and the water layer is used for next step.

Step 3

Experimental Procedure

To a 50 mL single neck RBF (0.13 m mol) E is taken in 5 mL water. D(0.13 m mol) in 15 ml water is added at RT and the resulting solution isstirred for 24 h at RT. White precipitate formed during the reaction.Reaction mixture centrifuged and precipitate is given water wash andthen lyophilized to get F as white powder.

Synthesis of IO-179_02

Step-1

Experimental Procedure

To a 50 mL single neck RBF phosphopropionic acid A (0.77 mmol, 119 mg)was taken in 5 mL dry THF. Cholesterol (200 mg, 0.52 mmol) and DCC (160mg, 0.77 mmol) were added at 0° C. and the resulting solution wasstirred for 16 h at RT. White precipitate formed during the reaction.White ppt separated by filtration; washed with 5 ml THF. Solventevaporated and washed with hexane to get the product as 150 mg of whitepowder. ¹H NMR (CDCl₃): 0.67-2.66 (m), 4.22 (s), 5.36 (s), 8.18 (br, s).¹³C NMR (CDCl₃): 11.83, 18.74, 19.23, 21.04, 22.55, 22.81, 23.97, 24.28,24.71, 27.43, 28.00, 28.24, 29.84, 31.82, 31.94, 33.32, 35.85, 36.21,36.39, 36.96, 39.49, 39.73, 40.13, 42.31, 49.96, 56.23, 56.67, 122.88,139.48, 176.78 (d) ESIMS (−ve mode): 521.3 (M−H).

Step-2

Experimental Procedure

To a 25 mL single neck RBF A (69 mg, 0.13 mmol) was taken in 1 mL THF. B(15 mg, 0.26 mmol) in 1 ml water was added at 0° C. Immediate pptappeared. 2 ml water added to dissolve the ppt and the resultingsolution was stirred for 2 h at RT. Reaction mixture was givenchloroform wash and the water layer was used for next step.

Step 3

Experimental Procedure

To a 100 mL single neck RBF B (0.13 m mol) was taken in 5 mL water. A(0.13 m mol) in 15 ml water was added at RT and the resulting solutionwas stirred for 2 h at RT. White precipitate formed during the reaction.Reaction mixture centrifuged and precipitate was given water wash andthen lyophilized to get 50 mg of white powder.

Synthesis of IO-179_03

Experimental Procedure

Compound A is prepared according to the procedure described in thepreparation of IO-183_01. All the successive steps have carried outaccording to the preparation of IO-179_02.

Synthesis of IO-180_01

Experimental Procedure

To a 50 mL single neck RBF A (same compound as 60b step 1 product) (1mmol) was taken in 25 mL dry THF. Ethyl glycolate (1 mmol) and DCC (1mmol) and DMAP (0.1 mmol) were added at 0° C. and the resulting solutionwas stirred for 16 h at RT. Compound separated as pasty solid by silicagel column chromatography.

Experimental Procedure

To a 25 mL single neck RBF B (0.13 mmol) was taken in 3 mL THF. LiOH(0.26 mmol) in 1 ml water was added at 0° C. Reaction mixture wasstirred for 4 h at RT. Reaction mixture was given chloroform wash andthe water layer was used for next step.

Experimental Procedure

To a 100 mL single neck RBF D (0.13 m mol) was taken in 5 mL water. C(0.13 m mol) in 15 ml water was added dropwise at RT and the resultingsolution was stirred for 20 h at RT. White precipitate formed during thereaction. Reaction mixture centrifuged and precipitate was given waterwash and then lyophilized to get E as white powder.

Synthesis of IO-180_02

Experimental Procedure

To a 50 mL single neck RBF A (same compound as 60a step 1 product) (1mmol) is taken in 25 mL dry THF. Ethyl glycolate (1 mmol) and DCC (1mmol) and DMAP (0.1 mmol) are added at 0° C. and the resulting solutionis stirred for 16 h at RT. Compound is separated as pasty solid bysilica gel column chromatography.

Experimental Procedure

To a 25 mL single neck RBF B (0.13 mmol) is taken in 3 mL THF. LiOH(0.26 mmol) in 1 ml water is added at 0° C. Reaction mixture is stirredfor 4 h at RT. Reaction mixture is given chloroform wash and the waterlayer is used for next step.

Experimental Procedure

To a 100 mL single neck RBF D (0.13 m mol) is taken in 5 mL water. C(0.13 m mol) in 15 ml water is added at RT and the resulting solution isstirred for 20 h at RT. White precipitate formed during the reaction.Reaction mixture centrifuged and precipitate is given water wash andthen lyophilized to get E as white powder.

Synthesis of IO-180_03

Experimental Procedure

Compound E is prepared following the similar procedure described for thepreparation of IO-180_01.

Synthesis of IO-184_01

Step 1:

Experimental Procedure

To a 100 mL single neck RBF ester A (1.272 g, 2.27 mmol) was taken in 20mL of THF/H₂O (3:1) and cooled to 0° C. under ice bath. To this icecooled solution LiOH (136 mg, 5.67 mmol) was added and was stirred at rtfor overnight, the TLC was checked. After completion the reactionmixture was extracted with ethyl acetate and washed with sodiumdihydrogen sulphate solution (40 mL) and brine (20 mL) successively. Theorganic layer was dried over anhydrous Na₂SO₄ and concentrated undervacuum and column was performed to yield 1 gm of pure B as white powder.

Step 2:

Experimental Procedure

A mixture of B (532 mg, 1 m mol) and H₃PO₃ (2 mmol) is heated to 60° C.under N₂, until a homogeneous mixture is achieved PCl₃ (1 m mol) isadded dropwise and stirred at 60° C. for 2 h. The resulting mixture iscooled to room temperature and extracted by water. Water solution islyophilized to get compound C.

Experimental Procedure

To a 25 mL single neck RBF C (0.13 mmol) is taken in 1 mL THF. Sodiumhydroxide (0.54 mmol) in 2 ml water is added at 0° C. Resulting solutionis stirred for 2 h at RT. Reaction mixture has given chloroform wash andthe water layer is used for next step.

Experimental Procedure

To a 100 mL single neck RBF Aquated platinum diaminocyclohexane (0.13 mmol) in 15 ml water is taken. D (0.13 m mol) in 5 mL water is added atRT and the resulting solution is stirred for 2 h at RT. Whiteprecipitate formed during the reaction. Reaction mixture centrifuged andprecipitate was given water wash to get E as white powder.

Synthesis of IO-190_01

8-Hydroxyquinoline (7.34 g, 0.05 mol) was dissolved in a continuouslystirred solution of 66.7 mL of distilled water and 3 mL of concentratedsulfuric acid at 15-18° C. Sodium nitrite (3.67 g) in distilled water(6.78 mL), was added drop wise to the reaction mixture over a period of30-40 min at 15-18° C., mixture was maintained at this temperature for 3h. The reaction mixture was neutralized with 40% sodium hydroxidesolution. It was then acidified with glacial acetic acid to pH 3.0-4.0.Yellow precipitate obtained was filtered, washed with distilled water,and dried. Yield: 6.7 g (89.5%).

0.174 g (0.01 mol) of 5-nitroso-8-hydroxyquinoline in 25 mL ofconcentrated hydrochloric acid was allowed to warm. To this was addedslowly, in small portions tin (Sn) metal (0.236 g, 0.02 mol). Thereaction mixture was heated at reflux for 6 h in boiling water bath. Thereaction mixture was allowed to cool to room temperature. To thereaction mixture was slowly added 20% solution of sodium hydroxide toget the precipitate. 5-Amino-8-hydroxyquinoline was extracted withether. Yield: 0.154 g (79.87%).

Cholesterol (1 g, 2.6 mmol) was dissolved in 40 mL of THF/DMF 1:1) and60% sodium hydride (w/w) in mineral oil (0.6 g, 15.5 mmol) was added,followed by stirring for 10 min. 2-bromo-1,1-dimethoxy ethane (1.21 mL,7.8 mmol) was added dropwise, and the mixture was stirred at 90° C.under reflux for 18 h. The mixture was cooled and CH₂Cl₂/MeOH (1:1) wasadded to eliminate excess NaH. After elimination of solvent waseliminated under vacuo, the residue was taken up in EtOAc, washedseveral times with water, dried over Na₂SO₄, filtered and concentrated.The crude was purified by flash column chromatography on silica gelusing 2-10% P.E. in EOAc, to obtain the product as a white solid, yield1.23 g 94%.

Trifluoroacetic acid/water (1:1) (2.5 mL 16.2 mmol) was added to asolution of cholesterol acetal (0.5 g 1 mmol) in 10 mL of CH₂Cl₂, andthe mixture was stirred at room temperature for 6 h. The mixture wasneutralized with 1N NaOH, extracted twice with CH₂Cl₂. dried overNa₂SO₄, filtered and concentrated, to obtain the product as white solid.

The product was obtained by refluxing stoichiometric amounts of thealdehyde (0.429 g, 1 mmol) and amine (0.160 g, 1 mmol) in absoluteethanol (15 ml) overnight in the presence of a catalytic amount oftrifluoroacetic acid. The desired product precipitated upon cooling thereaction mixture and it was subsequently purified by filtering andwashing with cold ethanol. Yield 0.4 g, 70%.

Sodium borohydride (0.875 g, 23.12 mmol) was added portion-wise tocholesterol quinoline (0.617 g, 1.08 mmol) in C₂H₅OH:THF mixture (1:1)at room temperature under inert atmosphere. After 6 hour stirring atroom temperature, the solvent was evaporated and the residue washed withsaturated brine solution and was extracted with DCM to get lightyellowish crystalline solid. Yield: 384 mg (62%).

To a 100 mL single neck RBF, cholesterol quinoline (0.151 g, 0.263 mmol)was taken in 3 mL of THF and cooled to 0° C. under ice bath. To this icecooled solution LiOH (11 mg, 0.263 mmol) in 1 mL H₂O was added and wasstirred at rt for 2 h, the TLC was checked. After completion thereaction mixture, THF was removed by rotavapour. Chloroform was added tothe reaction mixture. Compound was extracted with water. Then wholereaction mixture was used for the next reaction after rotavapourtreatment.

To a 50 mL single neck RBF, DACH platinum (100 mg, 0.263 mmol) was takenin 10 mL HPLC Water. To the above solution silver nitrate (89 mg, 0.526mmol) was added. The resulting solution was stirred under protectionfrom light at rt. After 24 h, AgCl precipitate was filtered. Filtratewas used for the next step.

To a 100 mL single neck RBF, Lithium salt of cholesterol quinoline (161mg, 0.263 mmol) was taken in 20 mL HPLC water and the resulting solutionwas stirred for 5 min at room temperature and to this solution DACH(OH₂)₂ platinum was added and it was stirred under protection from lightat rt for 24 h. The precipitate was filtered through filter paper andsimultaneously washed with HPLC water, HPLC Methanol and HPLC acetoneand dried. (Yield 60%).

Synthesis of Compound IO-185_01, IO-186_01, IO-187_01, IO-188_01,IO-189_01, IO-183_03, IO-183_04, IO-180_04

Synthesis of Aquated Cisplatin (B):

0.17 mmol diammineplatinum-dichloride (A), 0.34 mmol silver nitrate and7 mL water were added in a 25 mL RB and stirred for 48 h at room temp.Solution was centrifuged (4,000 rpm; 10 min) and white precipitate wasfiltered through syringe filter (25 mm/0.20 μm). Washed with 2 mL ofwater. Filtrate used for next reaction.

Synthesis of IO-185:

Procedure is similar as described for compound 25.

Synthesis of IO-186:

Procedure is similar as described for compound 26.

Synthesis of IO-187:

Procedure is similar as described for compound 27.

Synthesis of IO-188:

Procedure is similar as described for compound 28.

Synthesis of IO-189:

Procedure is similar as described for compound IO-131.

Synthesis of IO-183_03: Procedure is similar as described for compoundIO-183_01.

Synthesis of IO-183_04:

Procedure is similar as described for compound IO-183_02.

Synthesis of IO-180_04:

Procedure is similar as described for compound IO-180_01.

Example 8: Preparation of Lipid-Based Nanoparticles

Soy-phosphatidyl choline (fully hydrogenated, HSPC),1,2-Distearoyl-sn-Glycero-3-Phosphoethalonamine-N-[methoxy(Polyethyleneglycol)-2000] (Ammonium Salt) (DSPF-PFG-OMe) and cholesterol areselected as co-lipids. Liposomal nanoparticles are prepared bydissolving cholesterol-oxaliplatin lipid based platinum compounds of thepresent disclosure (as obtained in Examples 1 and 2) and colipids (HSPC,DSPF-PFG-OMe and cholesterol) in a 1:2:0.05:0.5 mol ratios respectively,in a mixture of dichloromethane and methanol in a glass vial. Theorganic solvent is removed with a gentle flow of moisture-free nitrogenand the remaining dried film of lipid is then kept under high vacuum forabout 8 hours. 300 mOsm buffer (sucrose and disodium hydrogenphosphate)is added to the vacuum-dried lipid film and the mixture is allowed tohydrate at 60° C. for 1 h. The vial is vortexed for about 2-3 minutes atroom temperature, and occasionally shaken in a 45° C. water bath toproduce multilamellar vesicles (MLV). Small unilamellar vesicles (SUV)are prepared bypassing of the MLV through extruder sequentially through400 μm, 200 μm and 100 μm membrane. The particle size of thenanoparticles obtained is measured by DLS instrument (Malvern).

Example 9: In-Vitro Cell Culture and Cell Viability Assays

The breast cancer cell line (4T1), cervical cancer cell line (HeLa) andLewis lung cancer cell line (LLC) are employed to study the cellviability assays. The 4T1 cells are cultured in RPMI1640 mediumsupplemented with 10% FBS, 50 unit ml-1 penicillin and 50 unit ml-1streptomycin-penicillin. HeLa and LLC cells are cultured in DMEM mediumsupplemented with 10% FBS, 50 unit ml-1 penicillin and 50 unit ml-1streptomycin-penicillin. The trypsinized cultured cancer cells areseeded into 96 well flat bottomed plates at a density of 3000 cells perwell one day prior to drug treatment. The following day, the platedcells are treated with various concentrations of nanoparticleformulations (as prepared by Example 3) with oxaliplatin as control. Theplates are then incubated for about 48 h in a 5% CO₂ atmosphere at about37° C. About 10 μl MTT reagent (10 mg/ml) is added and incubated for 2hours. The media is removed and the precipitate is solubilized in about100 μl of 1:1 DMSO-Methanol. The absorbance of the solubilizedprecipitate sample is measured in BioRad Elisa reader at 550 nm. Therelative cell viability is thereafter calculated from the recordedabsorption data.

The nanoparticle compositions of the present disclosure show significantcancer cell killing efficacy (FIG. 6). The said nanoparticles are testedin different cancer cell lines as described above and it is observedthat the compounds of the which have six-member co-ordination withplatinum (Compound 2, Compound 5) have a similar cell killing efficacywhen compared with oxaliplatin (control). Other compounds (Compound 1,Compound 6, Compound 3 and Compound 4) with five or seven memberplatinum co-ordination have better cell killing efficacy thanoxaliplatin. Most importantly, among these four compounds, Compound 4show significantly better cancer cell killing activity than oxaliplatincontrol.

In conclusion, the present disclosure aims at arriving at variousplatinum based amphiphiles as disclosed above. The said compounds have ageneral backbone of platinum-linker-lipid. Further, the disclosure alsorelates to carbenes, and more particularly, platinum containingcarbenes, wherein said platinum based carbenes are also employed as theplatinum moiety in platinum based amphiphiles. The various platinumbased amphiphiles of the present disclosure showcase significantlyimproved efficacy in cancer treatment and therefore, can be employed assuccessful alternatives in cancer treatment.

Example 10: Bioassays

Cell Culture:

Mammalian cells were grown in specific culture media, supplemented with10% fetal bovine serum (FBS) and antibiotics in a humidified environmentcontaining 5% CO₂ at 37° C.

Cell Viability Assay:

The effects of supramolecular platinum conjugates on the viability ofcancer cells were measured using MTT assay. Cells in 100 μlculture-media were plated in 96-well plates (3000-5000 cells/well) andallowed to adhere overnight in a humidified environment containing 5%CO₂ at 37° C. Fresh media (100 μL) containing different concentrationsof compounds were added to cells and incubated for 72 hours. Followingincubation, cell viability was determined using the MTT assay. The MTTassay measures cell viability through assessment of active mitochondrialdehydrogenase, which converts MTT into water-insoluble purple formazancrystals. Cell viability was plotted as dose-response curves using curvefitting.

The effects of compounds (IO-125, IO-126, IO-127, IO-128, IO-131 andIO-148) were evaluated in vitro in comparison with oxaliplatin in breastcancer (MDA-MB-231), ovarian cancer (SKOV-3), cervical cancer (HeLa) andcolorectal cancer (SW-620 and HCT-116) cell lines. The results showed asignificant inhibition of cell viability in 0-25 μM concentration rangefor all IO-compounds tested, showing a dose-response relationship (FIGS.7A-7F). The IC₅₀ values for the individual IO-compounds were lower thanoxaliplatin, revealing better efficacy for these compounds on humancancer cells.

Cellular Uptake of Platinum Compounds:

MDA-MB-231 cells (1×10⁶) were seeded in 2 ml media per well of a 6-wellplate and grown for 24 hrs. Required volume of compounds was added toachieve a final concentration of 50 μM platinum equivalent per well.Cells were incubated for 5 hrs after adding the compounds, then washedtwice with PBS, harvested by trypsinization, resuspended in PBS andcounted. Cells were centrifuged and pellets stored at −80° C. untilfurther processing. The cell pellets were digested with 100 μl nitricacid at 70° C. for 4 hours. Following digestion, the samples werediluted in 2% HCl and the quantity of accumulated platinum wasdetermined using Atomic absorption spectroscopy (AAS). The assay wasvalidated using a linear standard curve, generated from serial dilutionsof certified stock platinum standard and cellular uptake of platinum wasexpressed as ng of platinum per 1×10⁵ cells.

The results indicate that the uptake of cisplatin and oxaliplatin aresimilar in MDA-MB-231 cells, while all IO-compounds tested show higheruptake (˜7-20 fold) (FIG. 8). These results demonstrate that whenadministered at platinum equivalent concentrations, the uptake ofIO-compounds is significantly higher in comparison to cisplatin oroxaliplatin in cancer cells.

Measurement of Platinum in Mouse Tumors:

4T1 breast cancer cells were implanted subcutaneously in Balb/c mice onday 0. Tumor-bearing mice were treated with oxaliplatin and IO-127 at adose equivalent to 8 mg/kg of platinum (n=3) on day 9 post tumorimplantation. Approximately 40 mg of tumor was weighed, ground to finepowder in mortar and pestle with liquid nitrogen and digested overnightin nitric acid at 110° C. in ace high pressure tubes to achievehomogeneity. After acidification, each sample was diluted with 2% HCland analyzed by atomic absorption spectrophotometry (AAS) to measureabsorbance associated with platinum content. The assay was validatedusing a linear standard curve, generated from serial dilutions ofcertified stock platinum standard. The averaged platinum concentrationwas reported as ng of platinum per milligram of tissue.

As shown in FIG. 9, there was a significantly higher accumulation ofplatinum (as quantified per gram of tissue using atomic absorptionspectrophotometry) in tumors for the IO-127 treated mice as compared totumors harvested from mice dosed with an equivalent amount ofoxaliplatin. Our findings suggest that enhanced uptake of platinumassociated with IO-compounds could account for increased cellularkilling in tumors.

All patents and other publications identified in the specification andexamples are expressly incorporated herein by reference for allpurposes. These publications are provided solely for their disclosureprior to the filing date of the present application. Nothing in thisregard should be construed as an admission that the inventors are notentitled to antedate such disclosure by virtue of prior invention or forany other reason. All statements as to the date or representation as tothe contents of these documents is based on the information available tothe applicants and does not constitute any admission as to thecorrectness of the dates or contents of these documents.

Although preferred embodiments have been depicted and described indetail herein, it will be apparent to those skilled in the relevant artthat various modifications, additions, substitutions, and the like canbe made without departing from the spirit of the invention and these aretherefore considered to be within the scope of the invention as definedin the claims which follow. Further, to the extent not alreadyindicated, it will be understood by those of ordinary skill in the artthat any one of the various embodiments herein described and illustratedcan be further modified to incorporate features shown in any of theother embodiments disclosed herein.

What is claimed is:
 1. A compound comprising: (a) a platinum moiety; and (b) a lipid connected to said platinum moiety.
 2. The compound of claim 1, (i) wherein the compound comprises a carbonyl moiety; or (ii) wherein the platinum moiety comprises a platinum atom that is conjugated to said lipid via covalent bond, coordinate bond or a combination thereof; or (iii) further comprising at least one linker between the platinum moiety and the lipid.
 3. The compound of claim 1, wherein the carbonyl moiety is a carboxylic acid that is succinic acid, malonic acid, oxalic acid, keto acid, or a combination thereof
 4. The compound of claim 1, wherein the compound is of Formula (VIII): Q-linker-lipid  (VIII) wherein: Q is

X₃ is (CH₂)_(n), CH₂—NH and C₄H₈; X₄ is CO or —CH—CH₃; Z is a platinum containing compound, wherein the platinum forms a part of the ring; and n is 0, 1, or
 2. 5. The compound of claim 1, wherein the linker is: (i) —X—CH₂—X₂—X₁—, wherein: X is NH; X₁ is C(O)O, C(O)NH, O(CH₂)—O, NH, or O; X₂ is (CH₂)_(n) or C(O); and n is 0, 1, 2, 3, 4, or 5; (ii) —(CH₂)_(n)O—, —(CH₂)_(n)NHC(O)O—, —(CH₂)_(n)OC(O)NH—, —(CH₂)_(n)C(O)NH(CH₂)_(m)O—, —(CH₂)_(n)O(CH₂)_(m)O—, —(CH₂)_(n)OC(O)—, (CH₂)_(n)NHC(O)(CH₂)_(m)O—, or —(CH₂)_(n)C(O)O—; wherein, n and m are, independently, 0, 1, 2, 3, 4, or 5; (iii) —X₃—X₄X₅—X₆, wherein: X₃ is CH, CH₂, or O; and X₄, X₅ and X₆ are, independently, —CH₂O— or O; or (iv) a combination of (i)-(iii).
 6. The compound of claim 1, wherein the compound has (i) increased cellular uptake of platinum relative to cisplatin or oxaliplatin in cancer cells or the compound has a higher accumulation of platinum in a tumor relative to cisplatin or oxaliplatin at an equivalent dosage amount of amount of cisplatin or oxaliplatin; or (ii) a higher accumulation of platinum in a tumor relative to cisplatin or oxaliplatin at an equivalent dosage amount of amount of cisplatin or oxaliplatin.
 7. The compound of claim 1, that is of Formula (VIII): Q-linker-lipid  (VIII) wherein: Q is

wherein: X is NH or N(CH₂COO⁻); and Z is a platinum containing compound, wherein the platinum forms a part of the ring;

wherein: X is S⁺, C, S⁺═O, N⁺H, or P═O; X₁ is —CH, —CH₂ or —CH₂O; X₂ is C═O; and Z is a platinum containing compound, wherein the platinum forms a part of the ring;

wherein: X₁ is (CH₂)_(n); X₂ is C═O; Z is a platinum containing compound, wherein the platinum forms a part of the ring; and n is 0, 1, or 2;

wherein: R¹, R² and R³ are, independently, halogen, alkyl, amino, alkylamino, dialkylamino, hydroxyl, alkoxy, thiol, thioalkyl, O-acyl, -linker-lipid, or a combination thereof; or R₁ and R₂ together with the Pt atom or R₂ and R₃ together with the Pt atom form an optionally substituted cyclyl or heterocyclyl; or R₁ and R₂ together with the Pt atom and R₂ and R₃ together with the Pt atom form an optionally substituted cyclyl or heterocyclyl; or

wherein: R₁, R₂, R₃, R₄ and R₅ are, independently, halogen, alkyl, amino, alkylamino, dialkylamino, hydroxyl, alkoxy, thiol, thioalkyl, O-acyl, -linker-lipid, or a combination thereof; or R₁ and R₂ together with the Pt atom form an optionally substituted cyclyl or heterocyclyl; or R₃ and R₄ together with the Pt atom form an optionally substituted cyclyl or heterocyclyl; and the linker is one of (a)-(d): (a) —X—CH₂—X₂—X₁—, wherein: X is NH; X₁ is C(O)O, C(O)NH, O(CH₂)O, NH, or O; X₂ is (CH₂)_(n) or C(O); and n is 0, 1, 2, 3, 4, or 5; (b) —(CH₂)_(n)O—, —(CH₂)_(n)NHC(O)O—, —(CH₂)_(n)OC(O)NH—, —(CH₂)_(n)C(O)NH(CH₂)_(m)O—, —(CH₂)_(n)O(CH₂)_(m)O—, —(CH₂)_(n)OC(O)—, —(CH₂)_(n)NHC(O)(CH₂)_(m)O—, or —(CH₂)_(n)C(O)O—; wherein n and m are, independently, 0, 1, 2, 3, 4, or 5; (c) —X₃—X₄X₅—X₆—, wherein: X₃ is CH, CH₂, or O; and X₄, X₅ and X₆ are, independently, the same or different and are —CH₂O— or O; or (d) a combination of (i), (ii), or (iii).
 8. The compound of claim 1, that is of Formula (VIII): Q-linker-lipid  (VIII) wherein: Q is

wherein: X is NH or N(CH₂COO⁻); and Z is a platinum containing compound, wherein the platinum forms a part of the ring;

wherein: X is S⁺, C, S⁺═O, N⁺H, or P═O; X₁ is —CH, —CH₂ or —CH₂O; X₂ is C═O; and Z is a platinum containing compound, wherein the platinum forms a part of the rings

wherein: X₁ is (CH₂)_(n); X₂ is C═O; Z is a platinum containing compound, wherein the platinum forms a part of the ring; and n is 0, 1, or 2;

wherein: R¹, R² and R³ are, independently, halogen, alkyl, amino, alkylamino, dialkylamino, hydroxyl, alkoxy, thiol, thioalkyl, O-acyl, -linker-lipid, or a combination thereof; or R₁ and R₂ together with the Pt atom or R₂ and R₃ together with the Pt atom form an optionally substituted cyclyl or heterocyclyl; or R₁ and R₂ together with the Pt atom and R₂ and R₃ together with the Pt atom form an optionally substituted cyclyl or heterocyclyl; or

wherein: R₁, R₂, R₃, R₄ and R₅ are, independently, halogen, alkyl, amino, alkylamino, dialkylamino, hydroxyl, alkoxy, thiol, thioalkyl, O-acyl, -linker-lipid, or a combination thereof; or R₁ and R₂ together with the Pt atom form an optionally substituted cyclyl or heterocyclyl; or R₃ and R₄ together with the Pt atom form an optionally substituted cyclyl or heterocyclyl; and the linker is a bond, ethylene diamine, ethylene glycol, diethylene glycol, 1,3-propanediol, glycine, beta alanine, —O—, —CH₂ONHCH₂CH₂NHC(O)—, —NHCH₂CH₂NHC(O)O—, —NHCH₂CH₂—, —NHCH₂CH₂O—, —NHCH₂C(O)—, —NHCH₂C(O)O—, —NHCH₂C(O)OCH₂CH₂CH₂—, —NHCH₂C(O)OCH₂CH₂CH₂O—, —NHCH₂C(O)NH—, —CH₂CH₂—, —CH₂CH₂O—, —CH₂CH₂NHC(O)—, —CH₂CH₂NHC(O)O—, —CH₂CH₂O—, —CH₂C(O)NHCH₂CH₂—, —CH₂C(O)NHCH₂CH₂O—, —CH₂CH₂OCH₂CH₂—, —CH₂CH₂OCH₂CH₂O—, —CH₂C(O)—, —CH₂C(O)O—, —CH₂CH₂CH₂—, —CH₂CH₂CH₂O—, ═CH—CH═CH₂—, ═CH—CH═CHCH₂O—, —CH═CHCH₂—, —CH═CHCH₂O—, —OCH₂CH₂O—, —CH₂—, —CH₂O—, —NHC(O)CH₂—, —NHC(O)CH₂O—, —C(O)CH₂—, —C(O)CH₂O—, —OC(O)CH₂—, —OC(O)CH₂O—, —C(O)CH₂CH₂C(O)NHCH₂CH₂—, —OC(O)CH₂CH₂C(O)NHCH₂CH₂—, —C(O)CH₂CH₂C(O)NHCH₂CH₂O—, —OC(O)CH₂CH₂C(O)NHCH₂CH₂O—, —C(O)CH₂CH₂C(O)NHCH₂CH₂NHC(O)—, —OC(O)CH₂CH₂C(O)NHCH₂CH₂NHC(O)—, —C(O)CH₂CH₂C(O)NHCH₂CH₂NHC(O)O—, —OC(O)CH₂CH₂C(O)NHCH₂CH₂NHC(O)O—, or a combination thereof.
 9. A compound of Formula (V):

wherein: X₁, X₂, X₃ and X₄ are, independently, O, P, S, Se, N, C, O-A, O—B, DACH, halide, chelated dicarboxylato linkage group, non-chelated dicarboxylato linkage group, or a combination thereof; A and B are, independently, C, P, S, N, or a combination thereof; and X₄ is optional.
 10. A nanoparticle containing a compound of claim 1, optionally further comprising a co-lipid, stabilizer, or a combination thereof.
 11. The nanoparticle of claim 10, wherein ratio of the compound to co-lipid and/or stabilizer ranges from 99:1 to 1:99 (w/w), (mol/mol) or (vol/vol).
 12. The nanoparticle of claim 10, wherein the nanoparticle comprises (i) soy-phosphatidyl choline and 1,2-distearoyl-sn-glycero-3-phosphoethanolamine-N-[methoxy(polyethylene glycol)-2000] as co-lipids, and wherein the ratio of the compound and the co-lipids ranges from about 1:1:0.01 to about 1:4:3; or (ii) L-α-phosphatidylcholine, hydrogenated (soy) and 1,2-distearoyl-sn-glycero-3-phosphoethanolamine-N-[methoxy(polyethylene glycol)-2000] (ammonium salt) as co-lipids, and wherein the ratio of the compound and the co-lipids ranges from about 1:1:0.01 to about 1:4:3.
 13. The nanoparticle of claim 10, wherein the nanoparticle has (i) increased cellular uptake of platinum relative to cisplatin or oxaliplatin in cancer cells or the nanoparticle has a higher accumulation of platinum in a tumor relative to cisplatin or oxaliplatin at an equivalent dosage amount of amount of cisplatin or oxaliplatin; or (ii) a higher accumulation of platinum in a tumor relative to cisplatin or oxaliplatin at an equivalent dosage amount of amount of cisplatin or oxaliplatin.
 14. A nanoparticle containing a compound of claim
 10. 15. A pharmaceutical composition comprising a nanoparticle of claim 14 and an excipient.
 16. A pharmaceutical composition comprising a compound of claim 1 and an excipient.
 17. The pharmaceutical composition of claim 16, wherein the excipient is a granulating agent, binding agent, lubricating agent, disintegrating agent, sweetening agent, glidant, anti-adherent, anti-static agent, surfactant, anti-oxidant, gum, coating agent, coloring agent, flavouring agent, plasticizer, preservative, suspending agent, emulsifying agent, plant cellulosic material, spheronization agent, or a combination thereof.
 18. The pharmaceutical composition of claim 16, that is formulated into a dosage form that is an injectable, tablet, lyophilized powder, liposomal suspension, or a combination thereof.
 19. A method of treating or managing cancer in a subject, the method comprising administering a therapeutically effective amount of a compound of claim 1 to a subject in need thereof.
 20. The method of claim 19, wherein the cancer is breast, head and neck, ovarian, testicular, pancreatic, oral-esophageal, gastrointestinal, liver, gall bladder, lung, melanoma, skin, sarcoma, blood, brain, glioblastoma, tumor of neuroectodermal origin, or a combination thereof.
 21. A method for preparing a compound of claim 1, comprising conjugating the lipid with the platinum moiety to obtain said compound.
 22. A method for preparing the nanoparticle of claim 10, comprising reacting a platinum compound comprising platinum moiety and a lipid connected to said platinum moiety with a co-lipid in presence of solvent.
 23. A compound that is 